{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,4]],"date-time":"2026-11-04T00:02:11Z","timestamp":1793750531618,"version":"4.6.1"},"reference-count":26,"publisher":"F1000 Research Ltd","license":[{"start":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T00:00:00Z","timestamp":1690243200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004489","name":"Mitacs","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004489","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000971","name":"ALS Association","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000971","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100009017","name":"ALS Society of Canada","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100009017","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000406","name":"Motor Neurone Disease Association","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100000406","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research Foundation","doi-asserted-by":"crossref","award":["FDN154305"],"award-info":[{"award-number":["FDN154305"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Genome Quebec"},{"DOI":"10.13039\/100013061","name":"Ontario Genomics","doi-asserted-by":"crossref","award":["OGI-210"],"award-info":[{"award-number":["OGI-210"]}],"id":[{"id":"10.13039\/100013061","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100008762","name":"Genome Canada","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100008762","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["f1000research.com"],"crossmark-restriction":false},"short-container-title":["F1000Res"],"abstract":"<ns4:p>\n                    Charged multivesicular body protein 2B is a subunit of the endosomal sorting complex required for transport III (ESRCT-III), a complex implicated in the lysosomal degradation pathway and formation of multivesicular bodies. Mutations to the\n                    <ns4:italic>CHMP2B<\/ns4:italic>\n                    gene can result in abnormal protein aggregates in neurons and is therefore predicted to be associated in neurodegenerative diseases, including across the ALS-FTD spectrum.\u00a0Through our standardized experimental protocol which compares read-outs in knockout cell lines and isogenic parental controls, this study aims to enhance the reproducibility of research on this target by characterizing eight commercial antibodies against charged multivesicular body protein 2b using Western Blot, immunoprecipitation, and immunofluorescence. We identified many high-performing antibodies and encourage readers to use this report as a guide to select the most appropriate antibody for their specific needs.\n                  <\/ns4:p>","DOI":"10.12688\/f1000research.139755.1","type":"journal-article","created":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T11:43:32Z","timestamp":1690285412000},"page":"884","update-policy":"https:\/\/doi.org\/10.12688\/f1000research.crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["The identification of high-performing antibodies for Charged multivesicular body protein 2b for use in Western Blot, immunoprecipitation and immunofluorescence"],"prefix":"10.12688","volume":"12","author":[{"given":"Walaa","family":"Alshafie","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maryam","family":"Fotouhi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Riham","family":"Ayoubi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Irina","family":"Shlaifer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4125-3608","authenticated-orcid":false,"given":"Kathleen","family":"Southern","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter S.","family":"McPherson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5906-025X","authenticated-orcid":false,"given":"Carl","family":"Laflamme","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"name":"NeuroSGC\/YCharOS\/EDDU collaborative group","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2560","published-online":{"date-parts":[[2023,7,25]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"105144","DOI":"10.1016\/j.nbd.2020.105144","article-title":"Lessons learned from CHMP2B, implications for frontotemporal dementia and amyotrophic lateral sclerosis.","volume":"147","author":"C Ugbode","year":"2021","journal-title":"Neurobiol. Dis."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"979","DOI":"10.3389\/fpls.2018.00979","article-title":"Biogenesis and Function of Multivesicular Bodies in Plant Immunity.","volume":"9","author":"X Li","year":"2018","journal-title":"Front. Plant Sci."},{"key":"ref3","doi-asserted-by":"publisher","first-page":"519-547","DOI":"10.1146\/annurev.cellbio.23.090506.123319","article-title":"Biogenesis and function of multivesicular bodies.","volume":"23","author":"R Piper","year":"2007","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"1074-1077","DOI":"10.1212\/01.wnl.0000231510.89311.8b","article-title":"ALS phenotypes with mutations in CHMP2B (charged multivesicular body protein 2B).","volume":"67","author":"N Parkinson","year":"2006","journal-title":"Neurology."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"806-808","DOI":"10.1038\/ng1609","article-title":"Mutations in the endosomal ESCRTIII-complex subunit CHMP2B in frontotemporal dementia.","volume":"37","author":"G Skibinski","year":"2005","journal-title":"Nat. Genet."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"8","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":"ref7","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":"ref8","doi-asserted-by":"publisher","first-page":"3163-3168","DOI":"10.1002\/pmic.201400441","article-title":"Version 4.0 of PaxDb: Protein abundance data, integrated across model organisms, tissues, and cell-lines.","volume":"15","author":"M Wang","year":"2015","journal-title":"Proteomics."},{"key":"ref9","article-title":"DepMap 19Q3 Public ed.","year":"2019"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"172","DOI":"10.12688\/f1000research.130126.1","article-title":"The identification of high-performing antibodies for Moesin for use in Western Blot, immunoprecipitation, and immunofluorescence [version 1; peer review: awaiting peer review].","volume":"12","author":"W Alshafie","year":"2023","journal-title":"F1000Res."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"308","DOI":"10.12688\/f1000research.131333.1","article-title":"The identification of high-performing antibodies for transmembrane protein 106B (TMEM106B) for use in Western blot, immunoprecipitation, and immunofluorescence [version 1; peer review: awaiting peer review].","volume":"12","author":"R Ayoubi","year":"2023","journal-title":"F1000Res."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"324","DOI":"10.12688\/f1000research.132628.1","article-title":"The identification of high-performing antibodies for Sequestosome-1 for use in Western blot, immunoprecipitation and immunofluorescence [version 1; peer review: awaiting peer review].","volume":"12","author":"R Ayoubi","year":"2023","journal-title":"F1000Res."},{"key":"ref13","doi-asserted-by":"publisher","first-page":"348","DOI":"10.12688\/f1000research.132249.1","article-title":"The identification of high-performing antibodies for Profilin-1 for use in Western blot, immunoprecipitation and immunofluorescence [version 1; peer review: awaiting peer review].","volume":"12","author":"R Ayoubi","year":"2023","journal-title":"F1000Res."},{"key":"ref14","doi-asserted-by":"publisher","first-page":"355","DOI":"10.12688\/f1000research.131851.1","article-title":"The identification of high-preforming antibodies for Ubiquilin-2 for use in Western Blot, immunoprecipitation, and immunofluorescence [version 1; peer review: awaiting peer review].","volume":"12","author":"I McDowell","year":"2023","journal-title":"F1000Res."},{"key":"ref15","doi-asserted-by":"publisher","first-page":"376","DOI":"10.12688\/f1000research.133220.2","article-title":"The identification of high-performing antibodies for RNA-binding protein FUS for use in Western Blot, immunoprecipitation, and immunofluorescence [version 1; peer review: 1 approved with reservations].","volume":"12","author":"W Alshafie","year":"2023","journal-title":"F1000Res."},{"key":"ref16","doi-asserted-by":"publisher","first-page":"403","DOI":"10.12688\/f1000research.133479.1","article-title":"The identification of high-performing antibodies for Coiled-coil-helix-coiled-coil-helix domain containing protein 10 (CHCHD10) for use in Western Blot, immunoprecipitation and immunofluorescence [version 1; peer review: awaiting peer review].","volume":"12","author":"R Ayoubi","year":"2023","journal-title":"F1000Res."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"277","DOI":"10.12688\/f1000research.131852.2","article-title":"The identification of high-performing antibodies for TDP-43 for use in Western Blot, immunoprecipitation and immunofluorescence [version 1; peer review: awaiting peer review].","volume":"12","author":"D Worrall","year":"2023","journal-title":"F1000Res."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"452","DOI":"10.12688\/f1000research.133696.1","article-title":"The identification of high-performing antibodies for Vacuolar protein sorting-associated protein 35 (hVPS35) for use in Western Blot, immunoprecipitation and immunofluorescence [version 1; peer review: awaiting peer review].","volume":"12","author":"R Ayoubi","year":"2023","journal-title":"F1000Res."},{"key":"ref19","doi-asserted-by":"publisher","first-page":"391","DOI":"10.12688\/f1000research.132952.1","article-title":"The identification of high-performing antibodies for Superoxide dismutase [Cu-Zn] 1 (SOD1) for use in Western blot, immunoprecipitation, and immunofluorescence [version 1; peer review: awaiting peer review].","volume":"12","author":"R Ayoubi","year":"2023","journal-title":"F1000Res."},{"key":"ref20","doi-asserted-by":"publisher","DOI":"10.5281\/zenodo.5717498","article-title":"Antibody screening by Immunofluorescence.","author":"W Alshafie","year":"2021","journal-title":"Zenodo."},{"key":"ref21","doi-asserted-by":"publisher","DOI":"10.5281\/zenodo.6370501","author":"M Fotouhi","year":"2022","journal-title":"Zenodo."},{"key":"ref22","doi-asserted-by":"publisher","DOI":"10.5281\/zenodo.8139356","author":"K Southern","year":"2023","journal-title":"Zenodo."},{"key":"ref23","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":"2022","journal-title":"Nucleic Acids Res."},{"key":"ref24","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":"ref25","doi-asserted-by":"publisher","DOI":"10.5281\/zenodo.5717510","article-title":"Antibody Screening by Immunoblot.","author":"R Ayoubi","year":"2021","journal-title":"Zenodo."},{"key":"ref26","doi-asserted-by":"publisher","DOI":"10.5281\/zenodo.5717516","article-title":"Antibody screening by Immunoprecitation.","author":"R Ayoubi","year":"2021","journal-title":"Zenodo."}],"container-title":["F1000Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/f1000research.com\/articles\/12-884\/v1\/xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/12-884\/v1\/pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/f1000research.com\/articles\/12-884\/v1\/iparadigms","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,23]],"date-time":"2023-08-23T11:05:41Z","timestamp":1692788741000},"score":1,"resource":{"primary":{"URL":"https:\/\/f1000research.com\/articles\/12-884\/v1"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,25]]},"references-count":26,"URL":"https:\/\/doi.org\/10.12688\/f1000research.139755.1","relation":{"has-review":[{"id-type":"doi","id":"10.5256\/f1000research.153058.r190521","asserted-by":"subject"},{"id-type":"doi","id":"10.5256\/f1000research.153058.r196057","asserted-by":"subject"},{"id-type":"doi","id":"10.5256\/f1000research.153058.r196057","asserted-by":"object"},{"id-type":"doi","id":"10.5256\/f1000research.153058.r190521","asserted-by":"object"}]},"ISSN":["2046-1402"],"issn-type":[{"value":"2046-1402","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,7,25]]},"assertion":[{"value":"Indexed","URL":"https:\/\/f1000research.com\/articles\/12-884\/v1#article-reports","order":0,"name":"referee-status","label":"Referee status","group":{"name":"current-referee-status","label":"Current Referee Status"}},{"value":"10.5256\/f1000research.153058.r190521, Emma L. Clayton, UK Dementia Research Institute, King's College London, London, 08 Aug 2023, version 1, indexed","URL":"https:\/\/f1000research.com\/articles\/12-884\/v1#referee-response-190521","order":0,"name":"referee-response-190521","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"10.5256\/f1000research.153058.r196057, Xiao-Xin Yan, Department of Anatomy and Neurobiology, Central South University Xiangya School of Medicine, Changsha, Hunan, 23 Aug 2023, version 1, indexed","URL":"https:\/\/f1000research.com\/articles\/12-884\/v1#referee-response-196057","order":1,"name":"referee-response-196057","label":"Referee Report","group":{"name":"article-reports","label":"Article Reports"}},{"value":"This work was supported in part by the ALS-Reproducible Antibody Platform (ALS-RAP). ALS-RAP is a private-public partnership created by the ALS Association (USA), the Motor Neurone Disease Association (UK), and the ALS Society of Canada. The grant was from a Canadian Institutes of Health Research Foundation Grant (FDN154305) and by the Government of Canada through Genome Canada, Genome Quebec and Ontario Genomics (OGI-210). RA and WA were supported by a Mitacs fellowship.\u202f\u202f\nThe 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"}]}}