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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.

Authors :
Ayoubi R
Fotouhi M
Southern K
McPherson PS
Laflamme C
Source :
F1000Research [F1000Res] 2023 Oct 16; Vol. 12, pp. 452. Date of Electronic Publication: 2023 Oct 16 (Print Publication: 2023).
Publication Year :
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.<br />Competing Interests: Competing interests: For this project, the laboratory of Peter McPherson developed partnerships with high-quality antibody manufacturers and knockout cell line providers. The partners provide antibodies and knockout cell lines to the McPherson laboratory at no cost. These partners include: - Abcam- Aviva Systems Biology- Bio Techne- Cell Signalling Technology- Developmental Studies Hybridoma Bank- GeneTex- Horizon Discovery - Proteintech - Synaptic Systems -Thermo Fisher Scientific.<br /> (Copyright: © 2023 Ayoubi R et al.)

Details

Language :
English
ISSN :
2046-1402
Volume :
12
Database :
MEDLINE
Journal :
F1000Research
Publication Type :
Academic Journal
Accession number :
38434631
Full Text :
https://doi.org/10.12688/f1000research.133696.2