Dissemin is shutting down on January 1st, 2025

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F1000Research, F1000Research, (7), p. 1649, 2019

DOI: 10.12688/f1000research.15707.2

F1000Research, F1000Research, (7), p. 1649, 2018

DOI: 10.12688/f1000research.15707.1

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Comparative study of commercially available and homemade anti-VAMP7 antibodies using CRISPR/Cas9-depleted HeLa cells and VAMP7 knockout mice

Journal article published in 2018 by Agathe Verraes, Beatrice Cholley, Thierry Galli ORCID, Sebastien Nola ORCID
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

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Preprint: archiving forbidden
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Postprint: archiving forbidden
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Published version: archiving allowed
Data provided by SHERPA/RoMEO

Abstract

VAMP7 (vesicle-associated membrane protein) belongs to the intracellular membrane fusion SNARE (Soluble N-ethylmaleimide-sensitive factor attachment protein receptors) protein family. In this study, we used CRISPR/Cas9 genome editing technology to generate VAMP7 knockout (KO) human HeLa cells and mouse KO brain extracts in order to test the specificity and the background of a set of commercially available and homemade anti-VAMP7 antibodies. We propose a simple profiling method to analyze western blotting and use visual scoring for immunocytochemistry staining to determine the extent of the antibodies’ specificity. Thus, we were able to rank the performance of a set of available antibodies and further showed an optimized procedure for VAMP7 immunoprecipitation, which we validated using wild-type and KO mouse brain extracts.