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Identification of high-performing antibodies for Vacuolar protein sorting-associated protein 35 (hVPS35) for use in Western Blot, immunoprecipitation and immunofluorescence [version 2; peer review: 1 approved]

Authors :
Riham Ayoubi
Maryam Fotouhi
Kathleen Southern
Peter S. McPherson
Carl Laflamme
Author Affiliations :
<relatesTo>1</relatesTo>Department of Neurology and Neurosurgery, Structural Genomics Consortium, The Montreal Neurological Institute, McGill University, Montreal, Québec, H3A 2B4, Canada
Source :
F1000Research. 12:452
Publication Year :
2023
Publisher :
London, UK: F1000 Research Limited, 2023.

Abstract

Vacuolar protein sorting-associated protein 35 is a subunit of the retromer complex, a vital constituent of the endosomal protein sorting pathway. The D620N mutation in the VPS35 gene has been reported to be linked to type 17 Parkinson’s Disease progression, the exact molecular mechanism remains to be solved. The scientific community would benefit from the accessibility of validated and high-quality anti-hVPS35 antibodies. In this study, we characterized thirteen hVPS35 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. 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.

Details

ISSN :
20461402
Volume :
12
Database :
F1000Research
Journal :
F1000Research
Notes :
Revised Amendments from Version 1 In the introduction section of this revised article, we have incorporated additional comprehensive information about PARK17, along with an explanation of what a high-performing antibody is in each tested application. To the results and discussion section, we have included a description as to why the authors and YCharOS initiative does not score nor interpret antibody characterization data., , [version 2; peer review: 1 approved]
Publication Type :
Academic Journal
Accession number :
edsfor.10.12688.f1000research.133696.2
Document Type :
data-paper
Full Text :
https://doi.org/10.12688/f1000research.133696.2