A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence

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To enhance our understanding of STING1-associated disease, it is essential to make high-performing antibodies accessible to the scientific community. This study aims to improve reliability of STING1 research as we have characterized sixteen STING1 commercial antibodies for western blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. These studies are part of a larger, collaborative initiative seeking to address antibody reproducibility issues by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs." } { "@context": "http://schema.org", "@type": "BreadcrumbList", "itemListElement": [ { "@type": "ListItem", "position": "1", "item": { "@id": "https://f1000research.com/", "name": "Home" } }, { "@type": "ListItem", "position": "2", "item": { "@id": "https://f1000research.com/browse/articles", "name": "Browse" } }, { "@type": "ListItem", "position": "3", "item": { "@id": "https://f1000research.com/articles/13-1049", "name": "A guide to selecting high-performing antibodies for STING1 (Uniprot..." } } ] } Home Browse A guide to selecting high-performing antibodies for STING1 (Uniprot... ALL Metrics - Views Downloads Get PDF Get XML Cite How to cite this article Ruíz Moleón V, Alende C, Fotouhi M et al. A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.12688/f1000research.155929.2 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. Close Copy Citation Details Export Export Citation Sciwheel EndNote Ref. Manager Bibtex ProCite Sente EXPORT Select a format first Track Share ▬ ✚ Data Note Revised A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] Vera Ruíz Moleón https://orcid.org/0000-0003-3728-3158 1 , Charles Alende https://orcid.org/0009-0005-4611-6134 1 , Maryam Fotouhi 1 , [...] Riham Ayoubi 1 , Kathleen Southern https://orcid.org/0000-0002-4125-3608 1 , Carl Laflamme https://orcid.org/0000-0001-5906-025X 1 , NeuroSGC/YCharOS/EDDU collaborative group , ABIF consortium Vera Ruíz Moleón https://orcid.org/0000-0003-3728-3158 1 , Charles Alende https://orcid.org/0009-0005-4611-6134 1 , [...] Maryam Fotouhi 1 , Riham Ayoubi 1 , Kathleen Southern https://orcid.org/0000-0002-4125-3608 1 , Carl Laflamme https://orcid.org/0000-0001-5906-025X 1 , NeuroSGC/YCharOS/EDDU collaborative group , ABIF consortium PUBLISHED 02 Jan 2025 Author details Author details 1 Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, Canada Vera Ruíz Moleón Roles: Investigation, Validation Charles Alende Roles: Investigation, Validation Maryam Fotouhi Roles: Investigation Riham Ayoubi Roles: Formal Analysis, Methodology, Validation, Visualization, Writing – Review & Editing Kathleen Southern Roles: Writing – Original Draft Preparation, Writing – Review & Editing Carl Laflamme Roles: Conceptualization, Formal Analysis, Funding Acquisition, Resources, Supervision, Validation, Writing – Review & Editing OPEN PEER REVIEW DETAILS REVIEWER STATUS This article is included in the YCharOS (Antibody Characterization through Open Science) gateway. Abstract Stimulator of interferon genes protein (STING1) is an immune adaptor protein which promotes innate immune defense mechanisms against pathogens. To enhance our understanding of STING1-associated disease, it is essential to make high-performing antibodies accessible to the scientific community. This study aims to improve reliability of STING1 research as we have characterized sixteen STING1 commercial antibodies for western blot, immunoprecipitation, and immunofluorescence using a standardized experimental protocol based on comparing read-outs in knockout cell lines and isogenic parental controls. These studies are part of a larger, collaborative initiative seeking to address antibody reproducibility issues by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs. READ ALL READ LESS Keywords Uniprot ID: Q86WV6, STING1, hSTING, MPYS, MITA, stimulator of interferon genes protein, mediator of IRF3 activation, transmembrane protein 173, antibody characterization, antibody validation, western blot, immunoprecipitation, immunofluorescence Corresponding Author(s) Carl Laflamme ( [email protected] ) Close Corresponding author: Carl Laflamme Competing interests: For this project, the laboratory of Peter McPherson developed partnerships with high-quality antibody manufacturers and KO cell line providers. The partners provide antibodies and KO cell lines to the McPherson laboratory at no cost. These partners include: - Abcam- ABCD antibodies-ABclonal- Aviva Systems Biology -Bio Techne -Cell Signalling Technology -Developmental Studies Hybridoma Bank -GeneTex – Horizon Discovery – Proteintech – Synaptic Systems –Thermo Fisher Scientific. Grant information: This work was supported by a grant from the Michael J. Fox Foundation for Parkinson’s Disease Research. It was also supported by the Government of Canada through Genome Canada, Genome Quebec, and Ontario Genomics (grant no. OGI-210). RA is supported by a Mitacs fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Copyright: © 2025 Ruíz Moleón V et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. How to cite: Ruíz Moleón V, Alende C, Fotouhi M et al. A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.12688/f1000research.155929.2 ) First published: 12 Sep 2024, 13 :1049 ( https://doi.org/10.12688/f1000research.155929.1 ) Latest published: 02 Jan 2025, 13 :1049 ( https://doi.org/10.12688/f1000research.155929.2 ) Revised Amendments from Version 1 The revised version includes updated formats for Figures 2 and 3 to enhance their clarity. Additionally, the molecular function of the STING1 protein has been described with greater precision in the introduction. The revised version includes updated formats for Figures 2 and 3 to enhance their clarity. Additionally, the molecular function of the STING1 protein has been described with greater precision in the introduction. See the authors' detailed response to the review by Jieya Shao READ REVIEWER RESPONSES Introduction Stimulator of interferon genes protein (STING1) is a conserved transmembrane protein involved in innate immune response mechanisms. 1 , 2 It is primarily activated by cyclic GMP-AMP (cGAMP), a second messenger generated by cGAS upon detection of cytosolic DNA from pathogens or damaged cells. 1 , 3 , 4 Upon activation, STING1 triggers downstream signaling pathways, including the phosphorylation of IRF3 by TBK1, leading to the production of type I interferons and pro-inflammatory cytokines. 5 , 6 This response plays a crucial role in antiviral defense, immune surveillance, and the regulation of inflammatory diseases. 5 , 6 This research is part of a broader collaborative initiative in which academics, funders and commercial antibody manufacturers are working together to address antibody reproducibility issues by characterizing commercial antibodies for human proteins using standardized protocols, and openly sharing the data. 7 – 9 Here we evaluated the performance of sixteen commercial antibodies for STING1 for use in western blot, immunoprecipitation, and immunofluorescence, enabling biochemical and cellular assessment of STING1 properties and function. The platform for antibody characterization used to carry out this study was endorsed by a committee of industry academic representatives. It consists of identifying human cell lines with adequate target protein expression and the development/contribution of equivalent knockout (KO) cell lines, followed by antibody characterization procedures using most commercially available antibodies against the corresponding protein. The standardized consensus antibody characterization protocols are openly available on Protocol Exchange, a preprint server (DOI: 10.21203/rs.3.pex-2607/v1 ). 10 The authors do not engage in result analysis or offer explicit antibody recommendations. Our primary aim is to deliver top-tier data to the scientific community, grounded in Open Science principles. This empowers experts to interpret the characterization data independently, enabling them to make informed choices regarding the most suitable antibodies for their specific experimental needs. Guidelines on how to interpret antibody characterization data found in this study are featured on the YCharOS gateway. 11 Results and discussion Our standard protocol involves comparing readouts from WT (wild type) and KO cells. 12 , 13 The first step was to identify a cell line(s) that expresses sufficient levels of a given protein to generate a measurable signal using antibodies. To this end, we examined the DepMap (Cancer Dependency Map Portal, RRID:SCR_017655) transcriptomics database to identify all cell lines that express the target at levels greater than 2.5 log 2 (transcripts per million “TPM” + 1), which we have found to be a suitable cut-off. 7 The THP-1 cell line expresses the STING1 transcript at 4.1 log 2 (TPM+1), which is above the average range of cancer cells analyzed. A STING1 KO in THP-1 was obtained from Abcam ( Table 1 ). THP-1 cells are small, round human leukemia monocytic cell line commonly used to study proteins involved in immune responses. Phorbol 12-myristate-13-acetate (PMA) treatment is required to differentiate THP-1 in suspension into macrophage-like adherent cells. 14 Table 1. Summary of the cell lines used. Institution Catalog number RRID (Cellosaurus) Cell line Genotype Abcam ab271147 CVCL_0006 THP-1 WT Abcam ab270493 CVCL_B1AK THP-1 STING1 KO For western blot experiments, WT and STING1 KO protein lysates were first separated on SDS-PAGE, transferred onto nitrocellulose membranes, and then probed with sixteen STING1 antibodies in parallel ( Table 2 , Figure 1 ). Table 2. Summary of the STING1 antibodies tested. Company Catalog number Lot number RRID (Antibody Registry) Clonality Clone ID Host Concentration (μg/μl) Vendors recommended applications Abcam ab181125 ** 1000056-10 AB_2916053 recombinant-mono EPR13130 rabbit 0.30 Wb, IF Abcam ab227704 ** 1004954-2 AB_3083474 recombinant-mono SP338 rabbit 1.50 Wb, IF Abcam ab227705 ** 1062328-1 AB_3076486 recombinant-mono SP339 rabbit 0.31 Wb Abcam ab239074 ** 1001652-16 AB_3068528 recombinant-mono EPR13130-55 rabbit 0.53 Wb, IP, IF ABclonal A21051 ** 6100001330 AB_3083450 recombinant-mono ARC57967 rabbit 0.29 Wb Cell Signaling Technology 13647 ** 9 AB_2732796 recombinant-mono D2P2F rabbit 0.06 Wb, IP Cell Signaling Technology 50494 ** 8 AB_2799375 recombinant-mono D1V5L rabbit 0.56 Wb, IP DSHB CPTC-STING1-1 ** 4/27/23-15ug/mL AB_3068346 recombinant-mono CPTC-STING1-1 rabbit 0.02 Wb, IF GeneTex GTX134373 43628 AB_2887262 polyclonal - rabbit 0.42 Wb, IF Novus Biologicals (Bio-Techne) NBP2-24683 G-4 AB_2868483 polyclonal - rabbit 1.00 Wb, IF Novus Biologicals (Bio-Techne) NBP3-18816 ** COMU01 AB_3068548 recombinant-mono 2922D rabbit 1.00 Wb, IF Proteintech 19851-1-AP 00118975 AB_10665370 polyclonal - rabbit 0.70 Wb, IP, IF R&D Systems (Bio-Techne) MAB7169 * CFWR0420041 AB_10971940 monoclonal 723505 mouse 0.20 Wb, IF Thermo Fisher Scientific 702993 ** 2842286 AB_2762404 recombinant-mono 2H1L5 rabbit 0.50 Wb Thermo Fisher Scientific MA5-26030 * YJ4087701 AB_2725437 monoclonal OTI4H1 mouse 1.00 Wb, IF Thermo Fisher Scientific MA5-32768 ** YJ4089249A AB_2810045 recombinant-mono JM03-47 rabbit 1.00 Wb * Monoclonal antibody. ** Recombinant antibody. Figure 1. STING1 antibody screening by western blot. Lysates from PMA-treated THP-1 WT and STING1 KO were prepared, and 40 μg of protein were processed for western blot with the indicated STING1 antibodies. The Ponceau stained transfers of each blot are presented to show equal loading of WT and KO lysates and protein transfer efficiency from polyacrylamide gels to the nitrocellulose membrane. Antibody dilutions were chosen according to the recommendations of the antibody supplier. Antibody dilution used: ab181125** at 1/1000, ab227704** at 1/200, ab227705** at 1/1000, ab239074** at 1/1000, A21051** at 1/1000, 13647** at 1/1000, 50494** at 1/1000, CPTC-STING1-1** at 1/10, GTX134373 at 1/1000, NBP2-24683 at 1/1000, NBP3-18816** at 1/1000, 19851-1-AP at 1/1000, MAB7169* at 1/1000, 702993** at 1/1000, MA5-26030* at 1/1000, MA5-32768** at 1/1000. Predicted band size: 42 kDa. *Monoclonal antibody, **Recombinant antibody. We then assessed the capability of all sixteen antibodies to capture STING1 from THP-1 protein extracts using immunoprecipitation techniques, followed by western blot analysis. For the immunoblot step, a specific STING1 antibody identified previously (refer to Figure 1 ) was selected. Equal amounts of the starting material (SM) and unbound fractions (UB), as well as the whole immunoprecipitate (IP) eluates were separated by SDS-PAGE ( Figure 2 ). Figure 2. STING1 antibody screening by immunoprecipitation. Lysates from PMA-treated THP-1 WT were prepared, and immunoprecipitation was performed using 1.0 mg of lysate and 2.0 μg of the indicated STING1 antibodies pre-coupled to Dynabeads protein A or protein G. Samples were washed and processed for western blot with the indicated STING1 antibody. For western blot, NBP3-18816** was used at 1/1000. The Ponceau stained transfers of each blot are shown. SM = 4% starting material; UB = 4% unbound fraction; IP = immunoprecipitate, HC = antibody heavy chain. *Monoclonal antibody, **Recombinant antibody. For immunofluorescence, sixteen antibodies were screened using a mosaic strategy. First, THP-1 WT and STING1 KO cells were labelled with different fluorescent dyes in order to distinguish the two cell lines, and the STING1 antibodies were evaluated. Both WT and KO lines imaged in the same field of view to reduce staining, imaging and image analysis bias ( Figure 3 ). Quantification of immunofluorescence intensity in hundreds of WT and KO cells was performed for each antibody tested, and the images presented in Figure 3 are representative of this analysis. 10 Figure 3. STING1 antibody screening by immunofluorescence. PMA-treated THP-1 WT and STING1 KO cells were labelled with a green or a far-red fluorescent dye, respectively. WT and KO cells were mixed and plated to a 1:1 ratio in a 96-well plate with optically clear flat-bottom. Cells were stained with the indicated STING1 antibodies and with the corresponding Alexa-fluor 555 coupled secondary antibody including DAPI. Acquisition of the blue (nucleus-DAPI), green (identification of WT cells), red (antibody staining) and far-red (identification of KO cells) channels was performed. Representative images of the merged blue and red (grayscale) channels are shown. WT and KO cells are outlined with green and magenta dashed line, respectively. When an antibody was recommended for immunofluorescence by the supplier, we tested it at the recommended dilution. The rest of the antibodies were tested at 1 and 2 μg/ml and the final concentration was selected based on the detection range of the microscope used and a quantitative analysis not shown here. Antibody dilution used: ab181125** at 1/300, ab227704** at 1/1000, ab227705** at 1/300, ab239074** at 1/500, A21051** at 1/300, 13647** at 1/60, 50494** at 1/500, CPTC-STING1-1** at 1/20, GTX134373 at 1/1000, NBP2-24683 at 1/1000, NBP3-18816** at 1/1000, 19851-1-AP at 1/200, MAB7169* at 1/200, 702993** at 1/250, MA5-26030* at 1/1000, MA5-32768** at 1/1000. Bars = 10 μm. *Monoclonal antibody, **Recombinant antibody. In conclusion, we have screened sixteen STING1 commercial antibodies by western blot, immunoprecipitation, and immunofluorescence by comparing the signal produced by the antibodies in human THP-1 WT and STING1 KO cells. To assist users in interpreting antibody performance, Table 3 outlines various scenarios in which antibodies may perform in all three applications. Several high-quality and renewable antibodies that successfully detect STING1 were identified in all applications. Researchers who wish to study STING1 in a different species are encouraged to select high-quality antibodies, based on the results of this study, and investigate the predicted species reactivity of the manufacturer before extending their research. Table 3. Illustrations to assess antibody performance in western blot, immunoprecipitation and immunofluorescence. Western blot Immunoprecipitation Immunofluorescence The underlying data for this study can be found on Zenodo, an open-access repository for which YCharOS has its own collection of antibody characterization reports. 15 , 16 Limitations Inherent limitations are associated with the antibody characterization platform used in this study. Firstly, the YCharOS project focuses on renewable (recombinant and monoclonal) antibodies and does not test all commercially available STING1 antibodies. YCharOS partners provide approximately 80% of all renewable antibodies, but some top-cited polyclonal antibodies may not be available through these partners. Secondly, the YCharOS effort employs a non-biased approach that is agnostic to the protein for which antibodies have been characterized. The aim is to provide objective data on antibody performance without preconceived notions about how antibodies should perform or the molecular weight that should be observed in western blot. As the authors are not experts in STING1, only a brief overview of the protein’s function and its relevance in disease is provided. STING1 experts are responsible for analyzing and interpreting observed banding patterns in western blots and subcellular localization in immunofluorescence. Thirdly, YCharOS experiments are not performed in replicates primarily due to the use of multiple antibodies targeting various epitopes. Once a specific antibody is identified, it validates the protein expression of the intended target in the selected cell line, confirms the lack of protein expression in the KO cell line and supports conclusions regarding the specificity of the other antibodies. All experiments are performed using master mixes, and meticulous attention is paid to sample preparation and experimental execution. In immunofluorescence, the use of two different concentrations serves to evaluate antibody specificity and can aid in assessing assay reliability. In instances where antibodies yield no signal, a repeat experiment is conducted following titration. Additionally, our independent data is performed subsequently to the antibody manufacturers internal validation process, therefore making our characterization process a repeat. Lastly, as comprehensive and standardized procedures are respected, any conclusions remain confined to the experimental conditions and cell line used for this study. The use of a single cell type for evaluating antibody performance poses as a limitation, as factors such as target protein abundance significantly impact results. 10 Additionally, the use of cancer cell lines containing gene mutations poses a potential challenge, as these mutations may be within the epitope coding sequence or other regions of the gene responsible for the intended target. Such alterations can impact the binding affinity of antibodies. This represents an inherent limitation of any approach that employs cancer cell lines. Methods The standardized protocols used to carry out this KO cell line-based antibody characterization platform was established and approved by a collaborative group of academics, industry researchers and antibody manufacturers. The detailed materials and step-by-step protocols used to characterize antibodies in western blot, immunoprecipitation and immunofluorescence are openly available on Protocol Exchange, a preprint server (DOI: 10.21203/rs.3.pex-2607/v1 ). 10 Brief descriptions of the experimental setup used to carry out this study can be found below. Cell lines and antibodies used Cell lines used and primary antibodies tested in this study are listed in Tables 1 and 2 , respectively. To ensure that the cell lines and antibodies are cited properly and can be easily identified, we have included their corresponding Research Resource Identifiers, or RRID. 17 , 18 Cells were cultured in RPMI 1640 (Gibco, cat. number A1049101) containing 10% fetal bovine serum (Wisent, cat. number 080450), 0.05 mM 2-Mercaptoethanol (Gibco, cat. number 21985023, 2 mM L-glutamine (Wisent cat. number 609-065), 100 IU penicillin and 100 μg/ml streptomycin (Wisent cat. number 450201). THP-1 WT and STING1 KO cells were treated with 200 ng/ml PMA (Abcam, cat. number ab147465) for 2 days. 200 ng/ml of PMA was added to fresh medium in both day 1 and day 2. 14 Peroxidase-conjugated goat anti-rabbit and anti-mouse antibodies (Thermo Fisher Scientific, cat. number 65-6120 and 62-6520) and Peroxidase-conjugated Protein A (MilliporeSigma, cat. number P8651) were used in western blot and immunoprecipitation. Alexa-555-conjugated goat anti-rabbit and anti-mouse secondary antibodies (Thermo Fisher Scientific, cat. number A-21429 and A-21424) were used in immunofluorescence. Antibody screening by western blot PMA-treated THP-1 WT and STING1 KO cells (listed in Table 1 ) were collected in RIPA buffer (25mM Tris-HCl pH 7.6, 150mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% SDS) (Thermo Fisher Scientific, cat. number 89901) supplemented with 1x protease inhibitor cocktail mix (MilliporeSigma, cat. number P8340). Lysates were sonicated briefly and incubated for 30 min on ice. Lysates were spun at ~110,000 × g for 15 min at 4°C and equal protein aliquots of the supernatants were analyzed by SDS-PAGE and western blot. BLUelf prestained protein ladder (GeneDireX, cat. number PM008-0500) was used. Western blots were performed with precast midi 10% Bis-Tris polyacrylamide gels (Thermo Fisher Scientific, cat. number WG1201BOX) ran with MES SDS buffer (Thermo Fisher Scientific, cat. number NP000202), loaded in LDS sample buffer (Thermo Fisher Scientific, cat. number NP0008) with 1× sample reducing agent (Thermo Fisher Scientific, cat. number NP0009) and transferred on nitrocellulose membranes. Proteins on the blots were visualized with Ponceau S staining (Thermo Fisher Scientific, cat. number BP103-10) which is scanned to show together with individual western blot. Blots were blocked with 5% milk for 1 hr, and antibodies were incubated overnight at 4°C with 5% milk in TBS with 0.1% Tween 20 (TBST) (Cell Signaling Technology, cat. number 9997). Following three washes with TBST, the peroxidase conjugated secondary antibody was incubated at a dilution of ~0.2 μg/ml in TBST with 5% milk for 1 hr at room temperature followed by three washes with TBST. Membranes were incubated with Pierce ECL (Thermo Fisher Scientific, cat. number 32106) or Clarity Western ECL Substrate (Bio-Rad, cat. number 1705061) prior to detection with the iBright™ CL1500 Imaging System (Thermo Fisher Scientific, cat. number A44240). Antibody screening by immunoprecipitation Antibody-beads conjugates were prepared by adding 2 μg of antibody (with an exception 20 μl of antibody CPTC-STING1-1** and 20 μl of antibody 13647**) to 500 μl of Pierce IP Lysis Buffer (Thermo Fisher Scientific, cat. number 87788) in a microcentrifuge tube, together with 30μl of Dynabeads protein A- (for rabbit antibodies) or protein G- (for mouse antibodies) (Thermo Fisher Scientific, cat. number 10002D and 10004D, respectively). Tubes were rocked for ~1 hr at 4°C followed by two washes to remove unbound antibodies. PMA-treated THP-1 WT cells were collected in Pierce IP buffer (25 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40 and 5% glycerol) supplemented with protease inhibitors. Lysates were rocked for 30 min at 4°C and spun at 110,000 × g for 15 min at 4°C. 0.5 ml aliquots at 2.0 mg/ml of lysate were incubated with an antibody-bead conjugate for ~1 hr at 4°C. The unbound fractions were collected, and beads were subsequently washed three times with 1.0 ml of IP lysis buffer and processed for SDS-PAGE and western blot on precast midi 10% Bis-Tris polyacrylamide gels. Protein A:HRP (MilliporeSigma, cat. number P8651) was used as a secondary detection system at a concentration of 2.0 μg/ml. Antibody screening by immunofluorescence PMA-treated THP-1 WT and STING1 KO cells were labelled with a green and a far-red fluorescence dye, respectively (Thermo Fisher Scientific, cat. number C2925 and C34565, respectively). The nuclei were labelled with DAPI (Thermo Fisher Scientific, cat. Number D3571) fluorescent stain. WT and KO cells were plated in a 96-well plate with optically clear flat-bottom. (Perkin Elmer, cat. number 6055300) as a mosaic and incubated for 24 hrs in a cell culture incubator, 37 o C, 5% CO 2 . Cells were fixed in 4% paraformaldehyde (PFA) (Beantown chemical, cat. number 140770-10 ml) in phosphate buffered saline (PBS) (Wisent, cat. number 311-010-CL) for 15 min at room temperature and then washed 3 times with PBS. Cells were permeabilized in PBS with 0.1% Triton X-100 (Thermo Fisher Scientific, cat. number BP151-500) for 10 min at room temperature and blocked with PBS with 5% BSA, 5% goat serum (Gibco, cat. number 16210-064) and 0.01% Triton X-100 for 30 min at room temperature. Cells were incubated with IF buffer (PBS, 5% BSA, 0.01% Triton X-100) containing the primary STING1 antibodies overnight at 4°C. Cells were then washed 3 × 10 min with IF buffer and incubated with corresponding Alexa Fluor 555-conjugated secondary antibodies in IF buffer at a dilution of 1.0 μg/ml for 1 hr at room temperature with DAPI. Cells were washed 3 × 10 min with IF buffer and once with PBS. Images were acquired on an ImageXpress micro confocal high-content microscopy system (Molecular Devices), using a 20× NA 0.95 water immersion objective and scientific CMOS cameras, equipped with 395, 475, 555 and 635 nm solid state LED lights (lumencor Aura III light engine) and bandpass filters to excite DAPI, Cellmask Green, Alexa-555 and Cellmask Red, respectively. Images had pixel sizes of 0.68 × 0.68 microns, and a z-interval of 4 microns. For analysis and visualization, shading correction (shade only) was carried out for all images. Then, maximum intensity projections were generated using 3 z-slices. Segmentation was carried out separately on maximum intensity projections of Cellmask channels using CellPose 1.0, and masks were used to generate outlines and for intensity quantification. 19 Figures were assembled with Adobe Illustrator. Data availability Underlying data Zenodo: Antibody Characterization Report for STING1, https://doi.org/10.5281/zenodo.11582350 . 15 Zenodo: Dataset for the stimulator of interferon genes protein (STING1) antibody screening study, https://doi.org/10.5281/zenodo.12761294 . 16 Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0). Acknowledgment We would like to thank the NeuroSGC/YCharOS/EDDU collaborative group for their important contribution to the creation of an open scientific ecosystem of antibody manufacturers and KO cell line suppliers, for the development of community-agreed protocols, and for their shared ideas, resources, and collaboration. Members of the group can be found below. We would also like to thank the Advanced BioImaging Facility (ABIF) consortium for their image analysis pipeline development and conduction (RRID:SCR_017697). Members of each group can be found below. NeuroSGC/YCharOS/EDDU collaborative group: Thomas M. Durcan, Aled M. Edwards, Peter S. McPherson, Chetan Raina and Wolfgang Reintsch ABIF consortium: Claire M. Brown and Joel Ryan Thank you to the Structural Genomics Consortium, a registered charity (no. 1097737), for your support on this project. The Structural Genomics Consortium receives funding from Bayer AG, Boehringer Ingelheim, Bristol-Myers Squibb, Genentech, Genome Canada through Ontario Genomics Institute (grant no. OGI-196), the EU and EFPIA through the Innovative Medicines Initiative 2 Joint Undertaking (EUbOPEN grant no. 875510), Janssen, Merck KGaA (also known as EMD in Canada and the United States), Pfizer and Takeda. An earlier version of this of this article can be found on Zenodo (DOI: 10.5281/zenodo.11582350 ). References 1. Ishikawa H, Barber GN: STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling. Nature. 2008; 455 (7213): 674–678. PubMed Abstract | Publisher Full Text | Free Full Text 2. Ishikawa H, Ma Z, Barber GN: STING regulates intracellular DNA-mediated, type I interferon-dependent innate immunity. Nature. 2009; 461 (7265): 788–792. PubMed Abstract | Publisher Full Text | Free Full Text 3. Motwani M, Pesiridis S, Fitzgerald KA: DNA sensing by the cGAS-STING pathway in health and disease. Nat. Rev. Genet. 2019; 20 (11): 657–674. PubMed Abstract | Publisher Full Text 4. Zhang R, Kang R, Tang D: The STING1 network regulates autophagy and cell death. Signal Transduct. Target. Ther. 2021; 6 (1): 208. PubMed Abstract | Publisher Full Text | Free Full Text 5. Honda K, Takaoka A, Taniguchi T: Type I interferon [corrected] gene induction by the interferon regulatory factor family of transcription factors. Immunity. 2006; 25 (3): 349–360. PubMed Abstract | Publisher Full Text 6. Shinde O, Li P: The molecular mechanism of dsDNA sensing through the cGAS-STING pathway. Adv Immunol. 2024; 162 : 1–21. PubMed Abstract | Publisher Full Text 7. Ayoubi R, Ryan J, Biddle MS, et al. : Scaling of an antibody validation procedure enables quantification of antibody performance in major research applications. elife. 2023; 12 : 12. Publisher Full Text 8. Carter AJ, Kraemer O, Zwick M, et al. : Target 2035: probing the human proteome. Drug Discov. Today. 2019; 24 (11): 2111–2115. PubMed Abstract | Publisher Full Text 9. Licciardello MP, Workman P: The era of high-quality chemical probes. RSC Med. Chem. 2022; 13 (12): 1446–1459. PubMed Abstract | Publisher Full Text | Free Full Text 10. Ayoubi R, Ryan J, Bolivar SG, et al. : A consensus platform for antibody characterization (Version 1). Protocol Exchange. 2024. Publisher Full Text 11. Biddle MS, Virk HS: YCharOS open antibody characterisation data: Lessons learned and progress made. F1000Res. 2023; 12 (12): 1344. Publisher Full Text 12. Laflamme C, McKeever PM, Kumar R, et al. : Implementation of an antibody characterization procedure and application to the major ALS/FTD disease gene C9ORF72. elife. 2019; 8 : 8. Publisher Full Text 13. Alshafie W, Fotouhi M, Shlaifer I, et al. : Identification of highly specific antibodies for Serine/threonine-protein kinase TBK1 for use in immunoblot, immunoprecipitation and immunofluorescence. F1000Res. 2022; 11 : 977. Publisher Full Text 14. Starr T, Bauler TJ, Malik-Kale P, et al. : The phorbol 12-myristate-13-acetate differentiation protocol is critical to the interaction of THP-1 macrophages with Salmonella Typhimurium. PLoS One. 2018; 13 (3): e0193601. PubMed Abstract | Publisher Full Text | Free Full Text 15. Ruiz Moleon V, et al. : Antibody Characterization Report for STING1 (Uniprot ID: Q86WV6).2024. Publisher Full Text 16. Ayoubi R, Laflamme C: Dataset for the stimulator of interferon genes protein (STING1) antibody screening study. [Data set]. Zenodo 2024. Publisher Full Text 17. Bandrowski A, Pairish M, Eckmann P, et al. : The Antibody Registry: ten years of registering antibodies. Nucleic Acids Res. 2023; 51 (D1): D358–D367. PubMed Abstract | Publisher Full Text | Free Full Text 18. Bairoch A: The Cellosaurus, a Cell-Line Knowledge Resource. J. Biomol. Tech. 2018; 29 (2): 25–38. PubMed Abstract | Publisher Full Text | Free Full Text 19. Stringer C, Wang T, Michaelos M, et al. : Cellpose: a generalist algorithm for cellular segmentation. Nat. Methods. 2021; 18 (1): 100–106. PubMed Abstract | Publisher Full Text Comments on this article Comments (0) Version 2 VERSION 2 PUBLISHED 12 Sep 2024 ADD YOUR COMMENT Comment Author details Author details 1 Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, Canada Vera Ruíz Moleón Roles: Investigation, Validation Charles Alende Roles: Investigation, Validation Maryam Fotouhi Roles: Investigation Riham Ayoubi Roles: Formal Analysis, Methodology, Validation, Visualization, Writing – Review & Editing Kathleen Southern Roles: Writing – Original Draft Preparation, Writing – Review & Editing Carl Laflamme Roles: Conceptualization, Formal Analysis, Funding Acquisition, Resources, Supervision, Validation, Writing – Review & Editing Competing interests For this project, the laboratory of Peter McPherson developed partnerships with high-quality antibody manufacturers and KO cell line providers. The partners provide antibodies and KO cell lines to the McPherson laboratory at no cost. These partners include: - Abcam- ABCD antibodies-ABclonal- Aviva Systems Biology -Bio Techne -Cell Signalling Technology -Developmental Studies Hybridoma Bank -GeneTex – Horizon Discovery – Proteintech – Synaptic Systems –Thermo Fisher Scientific. Grant information This work was supported by a grant from the Michael J. Fox Foundation for Parkinson’s Disease Research. It was also supported by the Government of Canada through Genome Canada, Genome Quebec, and Ontario Genomics (grant no. OGI-210). RA is supported by a Mitacs fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Article Versions (2) version 2 Revised Published: 02 Jan 2025, 13:1049 https://doi.org/10.12688/f1000research.155929.2 version 1 Published: 12 Sep 2024, 13:1049 https://doi.org/10.12688/f1000research.155929.1 Copyright © 2025 Ruíz Moleón V et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Download Export To Sciwheel Bibtex EndNote ProCite Ref. Manager (RIS) Sente metrics Views Downloads F1000Research - - PubMed Central info_outline Data from PMC are received and updated monthly. - - Citations open_in_new 0 open_in_new 0 open_in_new SEE MORE DETAILS CITE how to cite this article Ruíz Moleón V, Alende C, Fotouhi M et al. A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.12688/f1000research.155929.2 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS track receive updates on this article Track an article to receive email alerts on any updates to this article. TRACK THIS ARTICLE Share Open Peer Review Current Reviewer Status: ? Key to Reviewer Statuses VIEW HIDE Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Version 2 VERSION 2 PUBLISHED 02 Jan 2025 Revised Views 0 Cite How to cite this report: Nielsen M and LaCava J. Reviewer Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.175918.r356517 ) The direct URL for this report is: https://f1000research.com/articles/13-1049/v2#referee-response-356517 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 05 Feb 2025 Mathias Nielsen , Laboratory of Cellular and Structural Biology, The Rockefeller University (Ringgold ID: 5929), New York, New York, USA John LaCava , University Medical Centre Groningen, The Rockefeller University, Groningen, The Netherlands Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.175918.r356517 The research presented by Moleón et al. is directed at demonstrating examples of standard operating procedures for the rigorous selection of antibodies to use in western blotting, immunoprecipitation, and immunofluorescence applications. The guidance given is sound and pertains to the ... Continue reading READ ALL The research presented by Moleón et al. is directed at demonstrating examples of standard operating procedures for the rigorous selection of antibodies to use in western blotting, immunoprecipitation, and immunofluorescence applications. The guidance given is sound and pertains to the principled selection of antibodies for these applications based on well-articulated criteria: the presence of signal in the WT and absence of signal in the cognate KO cell line (specificity in western); the depletion of signal in the unbound compared to the starting material along with the appearance of signal in the IP eluate (affinity to the target in IP), the presence of signal in the WT and absence of signal in the KO (specificity in IF). The authors are careful to state that the quality and pattern of the results are not necessarily transferable to other cell lines and experimental conditions, and are therefore representative of what one should expect from these reagents, given the experimental framework used. There are some intermediate or marginal cases presented within some of the data and various ways to interpret those. Some extended discussion about the meaning of such cases would be welcomed, but is not necessary to justify the work. Effectively, this article could help investigators looking for promising STING1 antibodies to short list pre-validated candidates, taken from this list, for screening in their applications, under their own experimental conditions - and evaluate those results in context, with the support of the comparative context provided here. It would have been satisfying to have mass spectrometry included in the analysis of the IP results - including the IP done in the WT vs KO for selected antibodies - or, at least to discuss a role for mass spectrometry in antibody validation for IP / co-IP (given that this is such a common application), but this appears to be outside of the intended scope of the research and not necessary to justify the work. This work, and work like it, contributes to more reproducible research by instructing investigators on principled ways to evaluate antibodies and, in this specific case, providing them with data that reduces their burden of evaluation for commercially available anti-STING1 products. I believe the following critiques apply and should be addressed: 1. Failure to indicate the units of the ingredients listed by %. Some will be e.g., by volume, such as 1% (v/v) NP-40, whereas others will be e.g., by weight, such as 0.1% (w/v) SDS - unlabeled % units appear through the methods for multiple reagents and it is best to ensure the reader is provided this info to complete the methods. 2. It seems that the experiments (WB, IP, and IF) are performed on PMA-treated (differentiated) THP-1 cells. The authors state that the PMA-treatment is required for differentiation into macrophage-like cells, but it is never explained why this treatment is relevant to the different experiments. The authors had already screened cell lines specifically for high STING1 expression and had chosen the THP-1 cell line - so why is the PMA-treatment needed? Is the expression of STING1 higher or lower in the differentiated cell? For IF experiments the treatment could be necessary, as the THP-1 cell line is a suspension cell that becomes adherent after the PMA treatment. So is the point to use the same input material / cell type for all experiments? One sentence could help clear this up. 3. In the Methods section under 'Antibody screening by western blot' the authors write: "Membranes were incubated with Pierce ECL (Thermo Fisher Scientific, cat. number 32106) or Clarity Western ECL Substrate (Bio-Rad, cat. number 1705061) prior to detection with the iBright™ CL1500 Imaging System (Thermo Fisher Scientific, cat. number A44240)." Why did the authors use two different ECL reagents? And does this affect the interpretation of the WB results? Since the results are presented as individual stand along panels - this should have no effect - but in the event that different panels are being compared, this would be a potential issues. The fact that two different reagents are used is not a problem, but it would be nice to add one line to the method or discussion stating that the blots are not intended to be compared to one another, per se, except through ratiometric means (e.g. UB/SM or IP/SM) and only qualitatively or semi-quantitatively (they mention these are only single replicate experiments, so, they would not be intended to be used quantitatively anyway). In essence, some verbiage on how these results can be used between panels (if at all), not just within panels, would be desirable. Is the rationale for creating the dataset(s) clearly described? Yes Are the protocols appropriate and is the work technically sound? Yes Are sufficient details of methods and materials provided to allow replication by others? Partly Are the datasets clearly presented in a useable and accessible format? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: Protein and nucleic acids biochemistry, proteomics / interactomics. We confirm that we have read this submission and believe that we have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however we have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Nielsen M and LaCava J. Reviewer Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.175918.r356517 ) The direct URL for this report is: https://f1000research.com/articles/13-1049/v2#referee-response-356517 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Tao Sc. Reviewer Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.175918.r356518 ) The direct URL for this report is: https://f1000research.com/articles/13-1049/v2#referee-response-356518 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 23 Jan 2025 Sheng-ce Tao , Shanghai Jiao Tong University, Shanghai, China Approved VIEWS 0 https://doi.org/10.5256/f1000research.175918.r356518 Because of the importance of STING, high quality antibody is a needed. This study compared sixteen commercially available STING1 antibodies for use in western blot, immunoprecipitation, and immunofluorescence. The antibodies were tested using standardized protocols in knockout cell lines and ... Continue reading READ ALL Because of the importance of STING, high quality antibody is a needed. This study compared sixteen commercially available STING1 antibodies for use in western blot, immunoprecipitation, and immunofluorescence. The antibodies were tested using standardized protocols in knockout cell lines and isogenic parental controls to assess their reliability. The aim of this study is helping researchers select the most suitable antibodies for perform STING related studies. Overall, I endorse the acceptance of this study. 1. What are the criteria for selecting the 16 commercial antibodies? Please clarify. 2. This study only tested one cell line (THP-1). It is not necessary that the conclusion from one cell generally also works among other cell lines. 3. Since one of the major application of these validated antibodies is WB, it is highly possible that these antibodies recognize linear epitopes on STING. In this case, mapping the epitopes (for example, using AbMap technology), and then judge the specificity by epitope searching across the whole proteome will be a more general approach. Is the rationale for creating the dataset(s) clearly described? Yes Are the protocols appropriate and is the work technically sound? Yes Are sufficient details of methods and materials provided to allow replication by others? Yes Are the datasets clearly presented in a useable and accessible format? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: Antibody technology, proteomics I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Tao Sc. Reviewer Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.175918.r356518 ) The direct URL for this report is: https://f1000research.com/articles/13-1049/v2#referee-response-356518 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Shao J. Reviewer Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.175918.r355436 ) The direct URL for this report is: https://f1000research.com/articles/13-1049/v2#referee-response-355436 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 13 Jan 2025 Jieya Shao , Washington University in St Louis, St. Louis, Missouri, USA Approved VIEWS 0 https://doi.org/10.5256/f1000research.175918.r355436 I approve ... Continue reading READ ALL I approve this revised report. Competing Interests: No competing interests were disclosed. Reviewer Expertise: molecular and cellular biology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Shao J. Reviewer Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.175918.r355436 ) The direct URL for this report is: https://f1000research.com/articles/13-1049/v2#referee-response-355436 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Version 1 VERSION 1 PUBLISHED 12 Sep 2024 Views 0 Cite How to cite this report: Shao J. Reviewer Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.171164.r333636 ) The direct URL for this report is: https://f1000research.com/articles/13-1049/v1#referee-response-333636 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 08 Nov 2024 Jieya Shao , Washington University in St Louis, St. Louis, Missouri, USA Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.171164.r333636 In this article the authors systematically compared the performance and specificity of sixteen commercial anti-STING1 antibodies using three different experimental approaches (Western blot, immunoprecipitation, and immunofluorescence staining). This study is part of a large collaborative initiative to characterize commercial antibodies ... Continue reading READ ALL In this article the authors systematically compared the performance and specificity of sixteen commercial anti-STING1 antibodies using three different experimental approaches (Western blot, immunoprecipitation, and immunofluorescence staining). This study is part of a large collaborative initiative to characterize commercial antibodies for different human proteins and offer unbiased guidance to researchers who can select the best antibodies for their specific experimental objectives based on their own interpretations. As a Data Note, this paper successfully achieved its goal with rigorously collected high-quality data, and the results are self-explanatory and useful for the research community. However, there are a few minor points to address. First, the authors wrote in the Introduction section that “activated in response to bacterial cyclic dinucleotides, STING1 initiates a cascade of signaling events”. This statement seems incorrect as cGAMP can be generated endogenously by cGAS rather than derived from bacteria. Second, the statement that aberrant expression of STING1 can lead to cancer needs clarification. Third, in Fig.2, the bottom panel of images are larger than the rest. Fourth, the images in Fig.3 can be rearranged (e.g. 4 x 4) for better presentation. Is the rationale for creating the dataset(s) clearly described? Yes Are the protocols appropriate and is the work technically sound? Yes Are sufficient details of methods and materials provided to allow replication by others? Yes Are the datasets clearly presented in a useable and accessible format? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: molecular and cellular biology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Shao J. Reviewer Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.171164.r333636 ) The direct URL for this report is: https://f1000research.com/articles/13-1049/v1#referee-response-333636 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 02 Jan 2025 Carl Laflamme , Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, H3A 2B4, Canada 02 Jan 2025 Author Response Dear Dr. Shao, Thank you for taking the time to review our STING1 antibody characterization report. We greatly appreciate your feedback. This report is part of a global ... Continue reading Dear Dr. Shao, Thank you for taking the time to review our STING1 antibody characterization report. We greatly appreciate your feedback. This report is part of a global initiative aimed at characterizing antibodies for every human protein. While the authors are experts in antibody development, we acknowledge that our expertise does not extend specifically to the STING1 protein. In light of your comments, we have revised the introduction to ensure greater accuracy and clarity regarding STING1. Additionally, we have modified Figures 2 and 3 in accordance with your recommendations. We trust these changes address your concerns and enhance the quality of our report. Thank you once again for your thoughtful insights. Best regards, Carl Dear Dr. Shao, Thank you for taking the time to review our STING1 antibody characterization report. We greatly appreciate your feedback. This report is part of a global initiative aimed at characterizing antibodies for every human protein. While the authors are experts in antibody development, we acknowledge that our expertise does not extend specifically to the STING1 protein. In light of your comments, we have revised the introduction to ensure greater accuracy and clarity regarding STING1. Additionally, we have modified Figures 2 and 3 in accordance with your recommendations. We trust these changes address your concerns and enhance the quality of our report. Thank you once again for your thoughtful insights. Best regards, Carl Competing Interests: No competing interests were disclosed. Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 02 Jan 2025 Carl Laflamme , Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, H3A 2B4, Canada 02 Jan 2025 Author Response Dear Dr. Shao, Thank you for taking the time to review our STING1 antibody characterization report. We greatly appreciate your feedback. This report is part of a global ... Continue reading Dear Dr. Shao, Thank you for taking the time to review our STING1 antibody characterization report. We greatly appreciate your feedback. This report is part of a global initiative aimed at characterizing antibodies for every human protein. While the authors are experts in antibody development, we acknowledge that our expertise does not extend specifically to the STING1 protein. In light of your comments, we have revised the introduction to ensure greater accuracy and clarity regarding STING1. Additionally, we have modified Figures 2 and 3 in accordance with your recommendations. We trust these changes address your concerns and enhance the quality of our report. Thank you once again for your thoughtful insights. Best regards, Carl Dear Dr. Shao, Thank you for taking the time to review our STING1 antibody characterization report. We greatly appreciate your feedback. This report is part of a global initiative aimed at characterizing antibodies for every human protein. While the authors are experts in antibody development, we acknowledge that our expertise does not extend specifically to the STING1 protein. In light of your comments, we have revised the introduction to ensure greater accuracy and clarity regarding STING1. Additionally, we have modified Figures 2 and 3 in accordance with your recommendations. We trust these changes address your concerns and enhance the quality of our report. Thank you once again for your thoughtful insights. Best regards, Carl Competing Interests: No competing interests were disclosed. Close Report a concern COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Bowman K. Reviewer Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.171164.r333635 ) The direct URL for this report is: https://f1000research.com/articles/13-1049/v1#referee-response-333635 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 07 Nov 2024 Karen Bowman , University of Leicester, Leicester, England, UK Approved VIEWS 0 https://doi.org/10.5256/f1000research.171164.r333635 The study involved characterisation of sixteen commercial antibodies, from nine different suppliers, for the stimulator of interferon genes protein (STING1) for use in western blot, immunoprecipitation, and immunofluorescence. The study formed part of the wider YCharOS collaboration to characterise commercial ... Continue reading READ ALL The study involved characterisation of sixteen commercial antibodies, from nine different suppliers, for the stimulator of interferon genes protein (STING1) for use in western blot, immunoprecipitation, and immunofluorescence. The study formed part of the wider YCharOS collaboration to characterise commercial antibodies for human proteins using standardized protocols with the intention of improving antibody reproducibility issues. As a Data Note, it allows the direct comparison of the performance of commercially available antibodies to STING1 protein to aid scientists in choosing the most appropriate antibody for their studies. This would be of use in several fields such as study of innate immunity, autoinflammatory diseases, cancer and neurodegenerative diseases. A committee of industry and academic representatives have endorsed the platform used, and the protocols used appear to be consistent with those in general use. The platform consisted of identification of a human cell line suitable for antibody characterisation studies i.e., with adequate levels of the STING1 protein to generate a measurable signal. They found that the commercially available THP-1 WT cell line expresses the STING1 transcript at a level above the average range of cancer cells analysed, and therefore, it was a logical choice for their study. The matched isogenic knockout control cell line, THP-1 STING1 KO was used as an appropriate negative control. The final step was a series of antibody characterisation procedures, limited to the most common research uses of antibodies. The interpretation of the results is left to the reader, with the study providing unbiased guidance. Limitations of the study are clearly stated. The summary Table 3 with illustrations to assess antibody performance in western blot, immunoprecipitation and immunofluorescence, is very helpful. A minor point to note. An ‘at a glance’ summary table encompassing the outcomes for each antibody would be useful. In conclusion, the study will be a useful resource when choosing the most appropriate antibody for study of the STING1 protein. Is the rationale for creating the dataset(s) clearly described? Yes Are the protocols appropriate and is the work technically sound? Yes Are sufficient details of methods and materials provided to allow replication by others? Yes Are the datasets clearly presented in a useable and accessible format? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: Cell biology. Drug discovery and development. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Bowman K. Reviewer Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.171164.r333635 ) The direct URL for this report is: https://f1000research.com/articles/13-1049/v1#referee-response-333635 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Comments on this article Comments (0) Version 2 VERSION 2 PUBLISHED 12 Sep 2024 ADD YOUR COMMENT Comment keyboard_arrow_left keyboard_arrow_right Open Peer Review Reviewer Status info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Reviewer Reports Invited Reviewers 1 2 3 4 Version 2 (revision) 02 Jan 25 read read read Version 1 12 Sep 24 read read Karen Bowman , University of Leicester, Leicester, UK Jieya Shao , Washington University in St Louis, St. Louis, USA Sheng-ce Tao , Shanghai Jiao Tong University, Shanghai, China Mathias Nielsen , The Rockefeller University (Ringgold ID: 5929), New York, USA John LaCava , University Medical Centre Groningen, The Rockefeller University, Groningen, The Netherlands Comments on this article All Comments (0) Add a comment Sign up for content alerts Sign Up You are now signed up to receive this alert Browse by related subjects keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 LaCava J et al. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 05 Feb 2025 | for Version 2 Mathias Nielsen , Laboratory of Cellular and Structural Biology, The Rockefeller University (Ringgold ID: 5929), New York, New York, USA John LaCava , University Medical Centre Groningen, The Rockefeller University, Groningen, The Netherlands 0 Views copyright © 2025 LaCava J et al. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The research presented by Moleón et al. is directed at demonstrating examples of standard operating procedures for the rigorous selection of antibodies to use in western blotting, immunoprecipitation, and immunofluorescence applications. The guidance given is sound and pertains to the principled selection of antibodies for these applications based on well-articulated criteria: the presence of signal in the WT and absence of signal in the cognate KO cell line (specificity in western); the depletion of signal in the unbound compared to the starting material along with the appearance of signal in the IP eluate (affinity to the target in IP), the presence of signal in the WT and absence of signal in the KO (specificity in IF). The authors are careful to state that the quality and pattern of the results are not necessarily transferable to other cell lines and experimental conditions, and are therefore representative of what one should expect from these reagents, given the experimental framework used. There are some intermediate or marginal cases presented within some of the data and various ways to interpret those. Some extended discussion about the meaning of such cases would be welcomed, but is not necessary to justify the work. Effectively, this article could help investigators looking for promising STING1 antibodies to short list pre-validated candidates, taken from this list, for screening in their applications, under their own experimental conditions - and evaluate those results in context, with the support of the comparative context provided here. It would have been satisfying to have mass spectrometry included in the analysis of the IP results - including the IP done in the WT vs KO for selected antibodies - or, at least to discuss a role for mass spectrometry in antibody validation for IP / co-IP (given that this is such a common application), but this appears to be outside of the intended scope of the research and not necessary to justify the work. This work, and work like it, contributes to more reproducible research by instructing investigators on principled ways to evaluate antibodies and, in this specific case, providing them with data that reduces their burden of evaluation for commercially available anti-STING1 products. I believe the following critiques apply and should be addressed: 1. Failure to indicate the units of the ingredients listed by %. Some will be e.g., by volume, such as 1% (v/v) NP-40, whereas others will be e.g., by weight, such as 0.1% (w/v) SDS - unlabeled % units appear through the methods for multiple reagents and it is best to ensure the reader is provided this info to complete the methods. 2. It seems that the experiments (WB, IP, and IF) are performed on PMA-treated (differentiated) THP-1 cells. The authors state that the PMA-treatment is required for differentiation into macrophage-like cells, but it is never explained why this treatment is relevant to the different experiments. The authors had already screened cell lines specifically for high STING1 expression and had chosen the THP-1 cell line - so why is the PMA-treatment needed? Is the expression of STING1 higher or lower in the differentiated cell? For IF experiments the treatment could be necessary, as the THP-1 cell line is a suspension cell that becomes adherent after the PMA treatment. So is the point to use the same input material / cell type for all experiments? One sentence could help clear this up. 3. In the Methods section under 'Antibody screening by western blot' the authors write: "Membranes were incubated with Pierce ECL (Thermo Fisher Scientific, cat. number 32106) or Clarity Western ECL Substrate (Bio-Rad, cat. number 1705061) prior to detection with the iBright™ CL1500 Imaging System (Thermo Fisher Scientific, cat. number A44240)." Why did the authors use two different ECL reagents? And does this affect the interpretation of the WB results? Since the results are presented as individual stand along panels - this should have no effect - but in the event that different panels are being compared, this would be a potential issues. The fact that two different reagents are used is not a problem, but it would be nice to add one line to the method or discussion stating that the blots are not intended to be compared to one another, per se, except through ratiometric means (e.g. UB/SM or IP/SM) and only qualitatively or semi-quantitatively (they mention these are only single replicate experiments, so, they would not be intended to be used quantitatively anyway). In essence, some verbiage on how these results can be used between panels (if at all), not just within panels, would be desirable. Is the rationale for creating the dataset(s) clearly described? Yes Are the protocols appropriate and is the work technically sound? Yes Are sufficient details of methods and materials provided to allow replication by others? Partly Are the datasets clearly presented in a useable and accessible format? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise Protein and nucleic acids biochemistry, proteomics / interactomics. We confirm that we have read this submission and believe that we have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however we have significant reservations, as outlined above. reply Respond to this report Responses (0) Nielsen M and LaCava J. Peer Review Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.175918.r356517) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/13-1049/v2#referee-response-356517 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Tao S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 23 Jan 2025 | for Version 2 Sheng-ce Tao , Shanghai Jiao Tong University, Shanghai, China 0 Views copyright © 2025 Tao S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Because of the importance of STING, high quality antibody is a needed. This study compared sixteen commercially available STING1 antibodies for use in western blot, immunoprecipitation, and immunofluorescence. The antibodies were tested using standardized protocols in knockout cell lines and isogenic parental controls to assess their reliability. The aim of this study is helping researchers select the most suitable antibodies for perform STING related studies. Overall, I endorse the acceptance of this study. 1. What are the criteria for selecting the 16 commercial antibodies? Please clarify. 2. This study only tested one cell line (THP-1). It is not necessary that the conclusion from one cell generally also works among other cell lines. 3. Since one of the major application of these validated antibodies is WB, it is highly possible that these antibodies recognize linear epitopes on STING. In this case, mapping the epitopes (for example, using AbMap technology), and then judge the specificity by epitope searching across the whole proteome will be a more general approach. Is the rationale for creating the dataset(s) clearly described? Yes Are the protocols appropriate and is the work technically sound? Yes Are sufficient details of methods and materials provided to allow replication by others? Yes Are the datasets clearly presented in a useable and accessible format? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise Antibody technology, proteomics I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (0) Tao Sc. Peer Review Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.175918.r356518) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/13-1049/v2#referee-response-356518 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Shao J. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 13 Jan 2025 | for Version 2 Jieya Shao , Washington University in St Louis, St. Louis, Missouri, USA 0 Views copyright © 2025 Shao J. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions I approve this revised report. Competing Interests No competing interests were disclosed. Reviewer Expertise molecular and cellular biology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (0) Shao J. Peer Review Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.175918.r355436) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/13-1049/v2#referee-response-355436 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2024 Shao J. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 08 Nov 2024 | for Version 1 Jieya Shao , Washington University in St Louis, St. Louis, Missouri, USA 0 Views copyright © 2024 Shao J. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions In this article the authors systematically compared the performance and specificity of sixteen commercial anti-STING1 antibodies using three different experimental approaches (Western blot, immunoprecipitation, and immunofluorescence staining). This study is part of a large collaborative initiative to characterize commercial antibodies for different human proteins and offer unbiased guidance to researchers who can select the best antibodies for their specific experimental objectives based on their own interpretations. As a Data Note, this paper successfully achieved its goal with rigorously collected high-quality data, and the results are self-explanatory and useful for the research community. However, there are a few minor points to address. First, the authors wrote in the Introduction section that “activated in response to bacterial cyclic dinucleotides, STING1 initiates a cascade of signaling events”. This statement seems incorrect as cGAMP can be generated endogenously by cGAS rather than derived from bacteria. Second, the statement that aberrant expression of STING1 can lead to cancer needs clarification. Third, in Fig.2, the bottom panel of images are larger than the rest. Fourth, the images in Fig.3 can be rearranged (e.g. 4 x 4) for better presentation. Is the rationale for creating the dataset(s) clearly described? Yes Are the protocols appropriate and is the work technically sound? Yes Are sufficient details of methods and materials provided to allow replication by others? Yes Are the datasets clearly presented in a useable and accessible format? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise molecular and cellular biology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (1) Author Response 02 Jan 2025 Carl Laflamme, Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, H3A 2B4, Canada Dear Dr. Shao, Thank you for taking the time to review our STING1 antibody characterization report. We greatly appreciate your feedback. This report is part of a global initiative aimed at characterizing antibodies for every human protein. While the authors are experts in antibody development, we acknowledge that our expertise does not extend specifically to the STING1 protein. In light of your comments, we have revised the introduction to ensure greater accuracy and clarity regarding STING1. Additionally, we have modified Figures 2 and 3 in accordance with your recommendations. We trust these changes address your concerns and enhance the quality of our report. Thank you once again for your thoughtful insights. Best regards, Carl View more View less Competing Interests No competing interests were disclosed. reply Respond Report a concern Shao J. Peer Review Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.171164.r333636) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/13-1049/v1#referee-response-333636 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2024 Bowman K. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 07 Nov 2024 | for Version 1 Karen Bowman , University of Leicester, Leicester, England, UK 0 Views copyright © 2024 Bowman K. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The study involved characterisation of sixteen commercial antibodies, from nine different suppliers, for the stimulator of interferon genes protein (STING1) for use in western blot, immunoprecipitation, and immunofluorescence. The study formed part of the wider YCharOS collaboration to characterise commercial antibodies for human proteins using standardized protocols with the intention of improving antibody reproducibility issues. As a Data Note, it allows the direct comparison of the performance of commercially available antibodies to STING1 protein to aid scientists in choosing the most appropriate antibody for their studies. This would be of use in several fields such as study of innate immunity, autoinflammatory diseases, cancer and neurodegenerative diseases. A committee of industry and academic representatives have endorsed the platform used, and the protocols used appear to be consistent with those in general use. The platform consisted of identification of a human cell line suitable for antibody characterisation studies i.e., with adequate levels of the STING1 protein to generate a measurable signal. They found that the commercially available THP-1 WT cell line expresses the STING1 transcript at a level above the average range of cancer cells analysed, and therefore, it was a logical choice for their study. The matched isogenic knockout control cell line, THP-1 STING1 KO was used as an appropriate negative control. The final step was a series of antibody characterisation procedures, limited to the most common research uses of antibodies. The interpretation of the results is left to the reader, with the study providing unbiased guidance. Limitations of the study are clearly stated. The summary Table 3 with illustrations to assess antibody performance in western blot, immunoprecipitation and immunofluorescence, is very helpful. A minor point to note. An ‘at a glance’ summary table encompassing the outcomes for each antibody would be useful. In conclusion, the study will be a useful resource when choosing the most appropriate antibody for study of the STING1 protein. Is the rationale for creating the dataset(s) clearly described? Yes Are the protocols appropriate and is the work technically sound? Yes Are sufficient details of methods and materials provided to allow replication by others? Yes Are the datasets clearly presented in a useable and accessible format? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise Cell biology. Drug discovery and development. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (0) Bowman K. Peer Review Report For: A guide to selecting high-performing antibodies for STING1 (Uniprot ID: Q86WV6) for use in western blot, immunoprecipitation, and immunofluorescence [version 2; peer review: 3 approved, 1 approved with reservations] . F1000Research 2025, 13 :1049 ( https://doi.org/10.5256/f1000research.171164.r333635) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/13-1049/v1#referee-response-333635 Alongside their report, reviewers assign a status to the article: Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. 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