{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T08:24:24Z","timestamp":1787214264361,"version":"build-2736575974"},"reference-count":13,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T00:00:00Z","timestamp":1781568000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"<ns5:p>TMEM175 is the pore-forming subunit of a lysosomal K+ channel complex that regulates lysosomal pH stability and membrane potential. To further investigate its cellular functions and implications in neurodegenerative diseases, antibody reagents are needed. Here we have characterized six TMEM175 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.<\/ns5:p>","DOI":"10.12688\/f1000research.182354.1","type":"journal-article","created":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T05:44:05Z","timestamp":1781588645000},"page":"942","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["A guide to selecting high-performing antibodies for TMEM175 (UniProt ID: Q9BSA9) for use in western blot, immunoprecipitation, and immunofluorescence"],"prefix":"10.12688","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3728-3158","authenticated-orcid":false,"given":"Vera Ru\u00edz","family":"Mole\u00f3n","sequence":"first","affiliation":[{"name":"Department of Neurology and Neurosurgery Montreal Quebec Structural Genomics Consortium, The Montreal Neurological Institute, McGill University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-4611-6134","authenticated-orcid":false,"given":"Charles","family":"Alende","sequence":"additional","affiliation":[{"name":"Department of Neurology and Neurosurgery Montreal Quebec Structural Genomics Consortium, The Montreal Neurological Institute, McGill University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maryam","family":"Fothouhi","sequence":"additional","affiliation":[{"name":"Department of Neurology and Neurosurgery Montreal Quebec Structural Genomics Consortium, The Montreal Neurological Institute, McGill University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4299-8281","authenticated-orcid":false,"given":"Sara Gonz\u00e1lez","family":"Bol\u00edvar","sequence":"additional","affiliation":[{"name":"Department of Neurology and Neurosurgery Montreal Quebec Structural Genomics Consortium, The Montreal Neurological Institute, McGill University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Riham","family":"Ayoubi","sequence":"additional","affiliation":[{"name":"Department of Neurology and Neurosurgery Montreal Quebec Structural Genomics Consortium, The Montreal Neurological Institute, McGill University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-7535-8718","authenticated-orcid":false,"given":"Vincent","family":"Francis","sequence":"additional","affiliation":[{"name":"Department of Neurology and Neurosurgery Montreal Quebec Structural Genomics Consortium, The Montreal Neurological Institute, McGill University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter S","family":"McPherson","sequence":"additional","affiliation":[{"name":"Department of Neurology and Neurosurgery Montreal Quebec Structural Genomics Consortium, The Montreal Neurological Institute, McGill University"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5906-025X","authenticated-orcid":false,"given":"Carl","family":"Laflamme","sequence":"additional","affiliation":[{"name":"Department of Neurology and Neurosurgery Montreal Quebec Structural Genomics Consortium, The Montreal Neurological Institute, McGill University"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2560","published-online":{"date-parts":[[2026,6,16]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"1101-1112","DOI":"10.1016\/j.cell.2015.08.002","article-title":"TMEM175 Is an Organelle K Channel Regulating Lysosomal Function.","volume":"162","author":"C Cang","year":"2015","journal-title":"Cell."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"431-437","DOI":"10.1038\/s41586-021-03185-z","article-title":"A growth-factor-activated lysosomal K+ channel regulates Parkinson's pathology.","volume":"591","author":"J Wie","year":"2021","journal-title":"Nature."},{"key":"ref3","doi-asserted-by":"publisher","first-page":"2292-2308.e20","DOI":"10.1016\/j.cell.2022.05.021","article-title":"Parkinson's disease-risk protein TMEM175 is a proton-activated proton channel in lysosomes.","volume":"185","author":"M Hu","year":"2022","journal-title":"Cell."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"4638-4640","DOI":"10.1111\/febs.16895","article-title":"Lysosome pH-regulated H\/K switching channel involved in maintaining lysosomal homeostasis.","volume":"290","author":"Y Wang","year":"2023","journal-title":"FEBS J."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"106244","DOI":"10.1016\/j.nbd.2023.106244","article-title":"TMEM175: A lysosomal ion channel associated with neurological diseases.","volume":"185","author":"L Wu","year":"2023","journal-title":"Neurobiol. Dis."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"1509-1545","DOI":"10.1038\/s41596-024-01095-8","article-title":"A consensus platform for antibody characterization.","volume":"20","author":"R Ayoubi","year":"2024","journal-title":"Nat. Protoc."},{"key":"ref7","doi-asserted-by":"crossref","first-page":"1344","DOI":"10.12688\/f1000research.141719.1","article-title":"YCharOS open antibody characterisation data: Lessons learned and progress made.","volume":"12","author":"M Biddle","year":"2023","journal-title":"F1000Res."},{"key":"ref8","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.48363","article-title":"Implementation of an antibody characterization procedure and application to the major ALS\/FTD disease gene C9ORF72.","volume":"8","author":"C Laflamme","year":"2019","journal-title":"elife."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"977","DOI":"10.12688\/f1000research.124632.1","article-title":"Identification of highly specific antibodies for Serine\/threonine-protein kinase TBK1 for use in immunoblot, immunoprecipitation and immunofluorescence.","volume":"11","author":"W Alshafie","year":"2022","journal-title":"F1000Res."},{"key":"ref10","doi-asserted-by":"publisher","first-page":"12","DOI":"10.7554\/eLife.91645.2","article-title":"Scaling of an antibody validation procedure enables quantification of antibody performance in major research applications.","volume":"12","author":"R Ayoubi","year":"2023","journal-title":"elife."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"D358-D367","DOI":"10.1093\/nar\/gkac927","article-title":"The Antibody Registry: ten years of registering antibodies.","volume":"51","author":"A Bandrowski","year":"2023","journal-title":"Nucleic Acids Res."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"25-38","DOI":"10.7171\/jbt.18-2902-002","article-title":"The Cellosaurus, a Cell-Line Knowledge Resource.","volume":"29","author":"A Bairoch","year":"2018","journal-title":"J. Biomol. Tech."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"100-106","DOI":"10.1038\/s41592-020-01018-x","article-title":"Cellpose: a generalist algorithm for cellular segmentation.","volume":"18","author":"C Stringer","year":"2021","journal-title":"Nat. Methods."}],"container-title":["F1000Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/f1000research.com\/articles\/15-942\/v1\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/15-942\/v1\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/15-942\/v1\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,20]],"date-time":"2026-08-20T07:49:10Z","timestamp":1787212150000},"score":1,"resource":{"primary":{"URL":"https:\/\/f1000research.com\/articles\/15-942\/v1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,16]]},"references-count":13,"URL":"https:\/\/doi.org\/10.12688\/f1000research.182354.1","relation":{"has-review":[{"id-type":"doi","id":"10.5256\/f1000research.201286.r495098","asserted-by":"subject"},{"id-type":"doi","id":"10.5256\/f1000research.201286.r506574","asserted-by":"subject"},{"id-type":"doi","id":"10.5256\/f1000research.201286.r495098","asserted-by":"object"},{"id-type":"doi","id":"10.5256\/f1000research.201286.r506574","asserted-by":"object"}]},"ISSN":["2046-1402"],"issn-type":[{"value":"2046-1402","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,16]]},"assertion":[{"value":"Indexed","URL":"https:\/\/f1000research.com\/articles\/15-942\/v1#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.5256\/f1000research.201286.r495098, Sian E. Evans, University of Leicester, Leicester, England, 17 Jul 2026, version 1, indexed","URL":"https:\/\/f1000research.com\/articles\/15-942\/v1#referee-response-495098","order":0,"name":"referee-response-495098","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.201286.r506574, Christian Tiede, University of Leeds, Leeds, England, 20 Aug 2026, version 1, indexed","URL":"https:\/\/f1000research.com\/articles\/15-942\/v1#referee-response-506574","order":1,"name":"referee-response-506574","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"This research was partly funded by the G-Can (GBA1-Canada) initiative, an open-science collaboration dedicated to advancing research on Parkinson's disease associated with GBA1 mutations. G-Can is supported by The Hilary & Galen Weston Foundation, Silverstein Foundation, and J. Sebastian van Berkom and Ghislaine Saucier. This work was also supported by the CQDM (by a grant from the Minist\u00e8re de l\u2019\u00c9conomie, de l\u2019Innovation et de l\u2019\u00c9nergie du Qu\u00e9bec) and 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.","order":2,"name":"grant-information","label":"Grant Information"},{"value":"This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.","order":0,"name":"copyright-info","label":"Copyright"}]}}