A guide to selecting high-performing antibodies for SCARB2 (UniProt ID: Q14108) for use in western blot, immunoprecipitation, and immunofluorescence

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McPherson" }, { "@type": "Person", "name": "Carl Laflamme" } ], "publisher": { "@type": "Organization", "name": "F1000Research", "logo": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 480, "width": 60 } }, "image": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 1200, "width": 150 }, "description": "Scavenger receptor class B member 2 (SCARB2), also known as lysosomal integral membrane protein type 2 (LIMP-2), is a multifunctional glycoprotein critical for lysosomal biogenesis and neuronal homeostasis. We characterized twelve commercially antibodies against SCARB2 in western blot, immunoprecipitation, and ten in immunofluorescence using a standardized workflow. Antibody performance was assessed by comparing signals in a wild-type cell line and its corresponding knockout derivative. This work is part of a broader collaborative public-good initiative to improve biomedical research by systematically evaluating commercial antibodies against human proteins and openly sharing the data as a resource for the scientific community. We encourage readers to use this report as a guide for selecting antibodies best suited to their specific applications." } { "@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/15-650", "name": "A guide to selecting high-performing antibodies for SCARB2 (UniProt..." } } ] } Home Browse A guide to selecting high-performing antibodies for SCARB2 (UniProt... ALL Metrics - Views Downloads Get PDF Get XML Cite How to cite this article Moleón VR, Alende C, Fothouhi M et al. A guide to selecting high-performing antibodies for SCARB2 (UniProt ID: Q14108) for use in western blot, immunoprecipitation, and immunofluorescence [version 1; peer review: awaiting peer review] . F1000Research 2026, 15 :650 ( https://doi.org/10.12688/f1000research.178929.1 ) 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 A guide to selecting high-performing antibodies for SCARB2 (UniProt ID: Q14108) for use in western blot, immunoprecipitation, and immunofluorescence [version 1; peer review: awaiting peer review] Vera Ruíz Moleón https://orcid.org/0000-0003-3728-3158 1 , Charles Alende https://orcid.org/0009-0005-4611-6134 1 , Maryam Fothouhi 1 , [...] Sara González Bolívar https://orcid.org/0000-0002-4299-8281 1 , Riham Ayoubi 1 , Vincent Francis https://orcid.org/0009-0000-7535-8718 1 , Peter S. McPherson 1 , Carl Laflamme https://orcid.org/0000-0001-5906-025X 1 Vera Ruíz Moleón https://orcid.org/0000-0003-3728-3158 1 , Charles Alende https://orcid.org/0009-0005-4611-6134 1 , [...] Maryam Fothouhi 1 , Sara González Bolívar https://orcid.org/0000-0002-4299-8281 1 , Riham Ayoubi 1 , Vincent Francis https://orcid.org/0009-0000-7535-8718 1 , Peter S. McPherson 1 , Carl Laflamme https://orcid.org/0000-0001-5906-025X 1 PUBLISHED 30 Apr 2026 Author details Author details 1 Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, Canada Vera Ruíz Moleón Roles: Investigation Charles Alende Roles: Investigation Maryam Fothouhi Roles: Investigation Sara González Bolívar Roles: Investigation Riham Ayoubi Roles: Supervision, Writing – Original Draft Preparation, Writing – Review & Editing Vincent Francis Roles: Writing – Original Draft Preparation, Writing – Review & Editing Peter S. McPherson Roles: Funding Acquisition, Supervision Carl Laflamme Roles: Conceptualization, Funding Acquisition, Writing – Review & Editing OPEN PEER REVIEW REVIEWER STATUS AWAITING PEER REVIEW This article is included in the YCharOS (Antibody Characterization through Open Science) gateway. Abstract Scavenger receptor class B member 2 (SCARB2), also known as lysosomal integral membrane protein type 2 (LIMP-2), is a multifunctional glycoprotein critical for lysosomal biogenesis and neuronal homeostasis. We characterized twelve commercially antibodies against SCARB2 in western blot, immunoprecipitation, and ten in immunofluorescence using a standardized workflow. Antibody performance was assessed by comparing signals in a wild-type cell line and its corresponding knockout derivative. This work is part of a broader collaborative public-good initiative to improve biomedical research by systematically evaluating commercial antibodies against human proteins and openly sharing the data as a resource for the scientific community. We encourage readers to use this report as a guide for selecting antibodies best suited to their specific applications. READ ALL READ LESS Keywords Q14108, SCARB2, SCARB2, 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 authors developed partnerships with leading antibody manufacturers and KO cell line providers. The partners provide antibodies and KO cell lines to this project at no cost. These partners include: Abcam, ABCD antibodies, ABclonal, Addgene, Aviva Systems Biology, BioTechne, Cell Signaling Technology, Developmental Studies Hybridoma Bank, GeneTex, Horizon Discovery, Institute for Protein Innovation, MilliporeSigma, Proteintech, Synaptic Systems, Thermo Fisher Scientific. Grant information: The author(s) declared that no grants were involved in supporting this work. Copyright: © 2026 Moleón VR 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: Moleón VR, Alende C, Fothouhi M et al. A guide to selecting high-performing antibodies for SCARB2 (UniProt ID: Q14108) for use in western blot, immunoprecipitation, and immunofluorescence [version 1; peer review: awaiting peer review] . F1000Research 2026, 15 :650 ( https://doi.org/10.12688/f1000research.178929.1 ) First published: 30 Apr 2026, 15 :650 ( https://doi.org/10.12688/f1000research.178929.1 ) Latest published: 30 Apr 2026, 15 :650 ( https://doi.org/10.12688/f1000research.178929.1 ) Introduction SCARB2 serves as a receptor for the lysosomal enzyme β-glucocerebrosidase (GCase), mediating its trafficking from the endoplasmic reticulum to lysosomes. 1 Dysfunction of this pathway contributes to the accumulation of glycolipids and lysosomal stress, mechanisms strongly implicated in neurodegenerative disorders such as Parkinson’s disease (PD) and action myoclonus–renal failure (AMRF) syndrome. 2 , 3 Here, we characterized twelve antibodies against SCARB2, for use in western blot, immunoprecipitation, and immunofluorescence using standardized protocols. Antibody performance was assessed by comparing signal in wild-type (WT) and knockout (KO) cell lines. The resulting data can guide the selection of antibodies best suited to specific research needs, enabling robust biochemical and cellular assessment of the target. This work is part of a broader collaborative initiative involving academics, funders, and antibody manufacturers, aimed at improving biomedical research reproducibility through the systematic characterization of commercial antibodies against human proteins and open data sharing. Antibodies are provided in-kind by participating manufacturers. The approach involves identifying cell lines with sufficient target expression, generating or sourcing corresponding KO cell lines, and evaluating commercially available antibodies, with a focus on renewable (monoclonal and recombinant) reagents. 4 We do not provide explicit antibody recommendations; rather, the data are presented to enable independent interpretation. Guidance on data interpretation is available via the YCharOS gateway 5 and in Table 5 of this data note. Results and discussion Antibody performance was assessed by comparing readouts from WT and KO cells. 4 The first step is to identify a cell line expressing sufficient levels of the target protein to generate a measurable signal using antibodies. To this end, we examined the DepMap transcriptomics database (Cancer Dependency Map Portal, RRID: SCR_017655 ) to identify cell lines with expression levels greater than 2.5 log 2 (transcripts per million “TPM” + 1), a threshold we have found to be suitable. 6 The MCF7 cell line expresses the SCARB2 transcript at 7.2 log 2 (TPM + 1), and a corresponding SCARB2 KO MCF7 cell line was obtained from Abcam ( Table 1 ). Moreover, as seen on DepMap, the MCF7 does not carry mutations in the SCARB2 gene that could affect antibody-epitope binding. Table 1. Summary of the cell lines used. Institution Catalog number RRID (Cellosaurus) Cell line Genotype Abcam ab271144 CVCL_0031 MCF7 WT Abcam ab274952 CVCL_B9BE MCF7 SCARB2 KO Primary antibodies are listed in Table 2 , with usage conditions summarized in Table 3 and secondary antibodies in Table 4 . Antibody usage is reported as dilution ratios to align with manufacturer recommendations and account for proprietary formulations. Table 2. Summary of the SCARB2 antibodies tested. Company Catalog number Lot number RRID (Antibody Registry) Clonality Clone ID Host Concentration (μg/μL) Vendors recommended applications Abcam ab176317 ** 1068975–2 AB_2620169 recombinant mono EPR12080 rabbit 0.10 Wb, IP Abcam ab196651 ** GR312647–3 AB_3101952 recombinant mono EPR12081 rabbit 0.84 Wb Abcam ab314217 ** 1065721-4 AB_3101784 recombinant mono EPR26243–125 rabbit 0.53 Wb, IP, IF ABclonal A9185 ** 4000001467 AB_2863681 recombinant mono ARC1467 rabbit 0.24 Wb Bio-Techne (R&D Systems) MAB1888 * JUZ024071 AB_2182970 monoclonal 220411 rat 1.00 Wb Bio-Techne (Novus Biologicals) NBP3–22186 ** 240465 AB_3097844 recombinant mono SR2211 rabbit NA Wb, IP Cell Signaling Technology 27960 ** 1 AB_3083079 recombinant mono E2Z5F rabbit 0.33 Wb, IP, IF Proteintech 27102–1-AP 00050639 AB_2880756 polyclonal - rabbit 0.50 Wb Thermo Fisher Scientific 702770 ** 2477085 AB_2723321 recombinant mono 12H5L1 rabbit 0.50 Wb Thermo Fisher Scientific 703037 ** 2942968 AB_2734813 recombinant mono 22H6L14 rabbit 0.50 Wb, IF Thermo Fisher Scientific 711805 ** 2384977 AB_2723322 recombinant poly 12HCLC rabbit 0.50 Wb Thermo Fisher Scientific 712072 ** 2854061 AB_2724602 recombinant poly - rabbit 0.50 Wb, IF ** Recombinant antibody. * Monoclonal antibody. Table 3. Dilutions of SCARB2 antibodies used in all western blot, immunoprecipitation and immunofluorescence. Catalog number Concentration (μg/μL) Vendor-recommended Wb dilution Wb dilution used IP volume (μL, for 2 μg input) Vendor-recommended IF dilution IF dilution tested (1) IF dilution tested (2) IF dilution shown ab176317 ** 0.10 1/1000–1/5000 1/1000 20.2 - 1/10 1/100 1/100 ab196651 ** 0.84 1/20000 1/20000 2.4 1/100 1/100 1/800 1/800 ab314217 ** 0.53 1/1000 1/1000 3.8 1/50 1/50 1/500 1/50 A9185 ** 0.24 1/500–1/1000 1/500 8.3 - 1/200 1/500 1/200 MAB1888 * 1.00 1/1000 1/1000 2.0 - - - - NBP3–22186 ** - 1/500–1/2000 1/500 5.0 - 1/200 1/500 1/200 27960 ** 0.33 1/1000 1/200 6.1 1/50–1/100 1/100 1/300 1/300 27102–1-AP 0.50 1/1000–1/8000 1/1000 4.0 - - - - 702770 ** 0.50 1/500 1/500 4.0 - 1/250 1/500 1/500 703037 ** 0.50 1/1000 1/1000 4.0 1/100 1/100 1/500 1/500 711805 ** 0.50 1/200 1/200 4.0 - 1/250 1/500 1/500 712072 ** 0.50 1/1000 1/1000 4.0 1/100 1/100 1/500 1/500 ** Recombinant antibody. * Monoclonal antibody. Table 4. Table of secondary antibodies used. Company Secondary antibody Catalog number RRID (Antibody Registry) Clonality Concentration (μg/μL) Working concentration (μg/mL) Proteintech HRP-Goat Anti-Rabbit Antibody (H + L) RGAR001 AB_3073505 recombinant polyclonal 1.0 0.05 Invitrogen HRP-Goat Anti-Rat Antibody (H + L) 31470 AB_228356 polyclonal 0.8 0.4 Cell Signaling Technology Protein A, HRP conjugate 12291 NA polyclonal 0.125 0.5 Proteintech CoraLite Plus 555-Goat Anti-Rabbit Antibody (H + L) RGAR003 AB_3073507 recombinant polyclonal 0.5 0.5 Invitrogen Alexa Fluor™ 555-Goat anti-Rat (H + L) A-21434 AB_141733 polyclonal 2 0.5 All twelve antibodies were first tested in western blot using WT and SCARB2 KO protein lysates ( Figure 1 ). This step enables concurrent validation of the KO cell line (absence of detectable SCARB2 signal) and identification of antibodies that detect SCARB2 with a band that is absent in the KO lane. Dilutions were selected based on supplier recommendations and adjusted when necessary ( Table 3 ). Figure 1. SCARB2 antibody screening by western blot. Lysates from MCF7 WT and SCARB2 KO were prepared and analyzed in western blot using the indicated SCARB2 antibodies. Ponceau-stained membranes are shown to confirm equal loading and efficient protein transfer from gel to membrane. Predicted molecular weight: 54 kDa. **Recombinant antibody; *Monoclonal antibody. The ability of each antibody to capture SCARB2 from MCF7 lysates was assessed by immunoprecipitation followed by western blot analysis ( Figure 2 ). Each immunoprecipitation was performed using 2 μg of antibody, or 5 μL when the concentration was not available ( Table 3 ). Starting material (SM), unbound (UB), and immunoprecipitated (IP) fractions were analyzed by SDS-PAGE and probed using antibody 702770 identified in the western blot screen. This assay evaluates target capture but does not assess selectivity (i.e., binding to SCARB2 versus off-target proteins). Figure 2. SCARB2 antibody screening by immunoprecipitation. MCF7 lysates were prepared, and immunoprecipitation was performed using the indicated SCARB2 antibodies. Immunoprecipitates were analyzed by western blot using SCARB2 antibody 702770** (1/500). Ponceau-stained membranes are shown. SM = 6% starting material; UB = 6% unbound fraction; IP = immunoprecipitate. ** = recombinant antibody; * = monoclonal antibody. For immunofluorescence, ten recombinant antibodies were screened at two dilutions using a mosaic strategy in which WT and KO cells, pre-labelled with distinct dyes, were imaged within the same field of view to minimize experimental bias ( Figure 3 ). Antibodies were tested at supplier-recommended dilutions when available; otherwise, they were tested at 1 and 2 μg/mL ( Table 3 ). Because detergent choice can strongly influence staining of lysosomal proteins, 7 both saponin and Triton X-100 were evaluated. Figure 3. SCARB2 antibody screening by immunofluorescence. MCF7 WT and SCARB2 KO cells were pre-labelled with green and far-red fluorescent dyes, respectively, mixed at a 1:1 ratio, and plated in 96-well plates. Cells were permeabilized with saponin or Triton X-100 and stained with the indicated SCARB2 antibodies. Representative images of the antibody and DAPI channels (grayscale) are shown. WT and KO cells are outlined with green and magenta dashed lines, respectively. WT/KO signal intensity ratios are indicated in the top-left corner of each antibody channel image. Scale bar = 10 μm. **Recombinant antibody; *Monoclonal antibody. Signal was quantified across at least 500 WT and KO cells, 4 and representative images are shown in Figure 3 . For each antibody, the WT/KO signal intensity ratio is reported: values near 1 indicate comparable signal in WT and KO cells, values >1 indicate enrichment in WT cells, and values <1 indicate higher signal in KO cells. The value shown corresponds to the condition yielding the highest ratio. These ratios facilitate comparison under the conditions tested but should be interpreted within the context of the specific assay and cell type. In conclusion, we screened twelve SCARB2 commercial antibodies in western blot, immunoprecipitation, and ten antibodies by immunofluorescence by comparing signal in WT and SCARB2 KO MCF7 cell lines. To assist with data interpretation, Table 5 outlines common antibody performance scenarios across applications. High-quality renewable antibodies detecting SCARB2 were identified for each application. Researchers studying SCARB2 in other species are encouraged to consider these results and verify predicted species reactivity with the manufacturer before proceeding. Table 5. Illustrations to assess antibody performance in all western blot, immunoprecipitation and immunofluorescence. Western blot Immunoprecipitation Immunofluorescence Limitations Inherent limitations are associated with the antibody characterization platform used in this study. First, the YCharOS project focuses on renewable (recombinant and monoclonal) antibodies and does not test all available antibodies against any given targets. While YCharOS partners provide access to the majority of renewable antibodies, some widely used polyclonal antibodies may not be included. Second, the YCharOS approach is protein-agnostic and aims to provide objective data on antibody performance without predefined expectations regarding molecular weight or localization. As such, only a brief overview of the protein target is provided, and expert interpretation of banding patterns and subcellular localization is encouraged. Third, experiments are not performed in biological replicates due to the parallel testing of multiple antibodies recognizing distinct epitopes. The identification of at least one specific antibody supports target expression in WT cells and its absence in KO cells, providing a reference for evaluating other antibodies. Experiments are performed using standardized conditions and master mixes to minimize variability. In immunofluorescence, testing at multiple antibody concentrations provides an additional assessment of specificity. Experiments may be repeated when no signal is detected. Furthermore, these data are generated independently of manufacturers’ validation processes, effectively constituting an external replication. Finally, conclusions are limited to the experimental conditions and cell line used. The reliance on a single cell type represents a constraint, as protein expression levels can influence antibody performance. The use of cancer cell lines also introduces potential confounders, including mutations that may affect antibody-epitope binding. These limitations are inherent to cell line-based validation approaches. Method The standardized protocols used for this antibody characterization study were established and approved by a collaborative group of academics, industry researchers, and antibody manufacturers. 4 Brief descriptions of the experimental procedures used in this study are described below. Cell lines The cell lines used in this study are listed in Table 1 . To facilitate proper citation and unambiguous identification, all cell lines and antibodies are referenced with their corresponding Research Resource Identifiers (RRIDs). 8 All cell lines used in this study were regularly tested for mycoplasma contamination and were confirmed to be mycoplasma-free. Primary and secondary antibodies Primary antibodies are listed in Table 2 with RRIDs. 9 Working dilutions for western blot and immunofluorescence, and volumes corresponding to 2 μg input for immunoprecipitation, are provided in Table 3 . Secondary antibodies and their working concentrations are listed in Table 4 . Antibody usage is reported as dilution ratios. Antibody screening by western blot MCF7 WT and SCARB2 KO cells were collected in RIPA buffer (25 mM Tris-HCl pH 7.6, 150 mM 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 30 min on ice. Lysates were spun at ~110,000 × g for 15 min at 4°C and 30 μg of 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 4-20% Tris-Glycine polyacrylamide gels (Thermo Fisher Scientific, cat. number WXP42012BOX) ran with Tris/Glycine/SDS buffer (Bio-Rad, cat. number 1610772), loaded in Laemmli loading sample buffer (Thermo Fisher Scientific, cat. number AAJ61337AD) 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 O/N at 4°C with 5% milk in TBS with 0.1% Tween 20 (TBST) (Cell Signalling 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) prior to detection with the iBright™ CL1500 Imaging System (Thermo Fisher Scientific, cat. number A44240). Antibody screening by immunoprecipitation Antibody-bead conjugates were prepared by adding 2 μg to 500 μl of Pierce IP Lysis Buffer from Thermo Fisher Scientific (cat. number 87788) in a microcentrifuge tube, together with 30 μl of Dynabeads protein A- (for rabbit antibodies) or protein G- (for rat antibodies) (Thermo Fisher Scientific, cat. number 10002D and 10004D, respectively). Tubes were rocked for ~1 h at 4°C followed by two washes to remove unbound antibodies. MCF7 WT 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 inhibitor. Lysates were rocked 30 min at 4°C and spun at 110,000 × g for 15 min at 4°C. 0.5 ml aliquots at 1 mg/ml of lysate were incubated with an antibody-bead conjugate for ~1 h at 4 °C. The unbound fractions were collected, and beads were subsequently washed three times with 1.0 ml of IP buffer and processed for SDS-PAGE and western blot on precast midi 4–20% Tris-Glycine polyacrylamide gels. Antibody screening by immunofluorescence MCF7 WT and SCARB2 KO cells were labelled with a green and a far-red fluorescence dye, respectively (Thermo Fisher Scientific, cat. number C2925 and C34565). The nuclei were labelled with DAPI (Thermo Fisher Scientific, cat. Number D3571) fluorescent stain. WT and KO cells were plated on 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 at 37°C, 5% CO 2 . Culture medium was removed, and cells were fixed in 4% paraformaldehyde (PFA) (VWR, cat. number 100503-917) in phosphate buffered saline (PBS) (Wisent, cat. number 311-010-CL) for 10 min at room temperature. Cells were permeabilized in PBS1x with 0.05% Saponin (MilliporeSigma, cat. number 47036) or 0.1% Triton X-100 (Thermo Fisher Scientific, cat. number BP151-500) for 10 min at room temperature. Cells were blocked in IF buffer (PBS1x with 5% BSA and 0.005% Saponin or 0.01% Triton X-100) with 5% goat serum (Gibco, cat. number 16210-064) for 1 h at room temperature. Cells were incubated with the corresponding IF buffer containing the primary SCARB2 antibodies overnight at 4°C. Cells were then washed 3 × 10 min with IF buffer and incubated with the corresponding Fluor 555-conjugated secondary antibody in IF buffer for 1 hr at room temperature. Cells were washed 3 × 10 min with PBS1x then incubated with DAPI and washed once with PBS1x. Images were acquired on an ImageXpress micro confocal high-content microscopy system (Molecular Devices), using a 20x NA 0.94 air 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. A minimum of 500 WT and KO cells were imaged per antibody. 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. 10 We have developed a collection of scripts in Python and in ImageJ/FIJII made openly available on GitHub ( https://github.com/ABIF-McGill/YCharOS_IF_characterization ). 4 Figures were assembled with Adobe Illustrator. Data availability Underlying data Dataset for the SCARB2 antibody screening study: doi.org/10.5281/zenodo.18805484 . 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. References 1. Gonzalez A, Valeiras M, Sidransky E, et al. :Lysosomal integral membrane protein-2: a new player in lysosome-related pathology. Mol Genet Metab. 2014; 111 (2): 84–91. PubMed Abstract | Publisher Full Text | Free Full Text 2. Rothaug M, Zunke F, Mazzulli JR, et al. :LIMP-2 expression is critical for β-glucocerebrosidase activity and α-synuclein clearance. P Natl Acad Sci USA. 2014; 111 (43): 15573–15578. Publisher Full Text 3. Berkovic SE, Dibbens LM, Oshlack A, et al. :Array-based gene discovery with three unrelated subjects shows SCARB2/LIMP-2 deficiency causes myoclonus epilepsy and glomerulosclerosis. Am J Hum Genet. 2008; 82 (3): 673–684. 4. Ayoubi R, Ryan J, Gonzalez Bolivar S, et al. :A consensus platform for antibody characterization. Nat Protoc. 2024; 20 : 1509–1545. Publisher Full Text 5. Biddle MS, Virk HS:YCharOS open antibody characterisation data: Lessons learned and progress made. F1000Res. 2023; 12 : 1344. PubMed Abstract | Publisher Full Text | Free Full Text 6. 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 7. Cuddy LK, Mazzulli JR:Analysis of lysosomal hydrolase trafficking and activity in human iPSC-derived neuronal models. STAR Protoc. 2021; 2 (1): 100340. PubMed Abstract | Publisher Full Text | Free Full Text 8. Bairoch A:The Cellosaurus, a Cell-Line Knowledge Resource. J Biomol Tech. 2018; 29 (2): 25–38. Publisher Full Text 9. 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 10. 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 1 VERSION 1 PUBLISHED 30 Apr 2026 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, Canada Vera Ruíz Moleón Roles: Investigation Charles Alende Roles: Investigation Maryam Fothouhi Roles: Investigation Sara González Bolívar Roles: Investigation Riham Ayoubi Roles: Supervision, Writing – Original Draft Preparation, Writing – Review & Editing Vincent Francis Roles: Writing – Original Draft Preparation, Writing – Review & Editing Peter S. McPherson Roles: Funding Acquisition, Supervision Carl Laflamme Roles: Conceptualization, Funding Acquisition, Writing – Review & Editing Competing interests For this project, the authors developed partnerships with leading antibody manufacturers and KO cell line providers. The partners provide antibodies and KO cell lines to this project at no cost. These partners include: Abcam, ABCD antibodies, ABclonal, Addgene, Aviva Systems Biology, BioTechne, Cell Signaling Technology, Developmental Studies Hybridoma Bank, GeneTex, Horizon Discovery, Institute for Protein Innovation, MilliporeSigma, Proteintech, Synaptic Systems, Thermo Fisher Scientific. Grant information The author(s) declared that no grants were involved in supporting this work. Article Versions (1) version 1 Published: 30 Apr 2026, 15:650 https://doi.org/10.12688/f1000research.178929.1 Copyright © 2026 Moleón VR 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. 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europepmc
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License: CC-BY-4.0