Identification of high-performing antibodies for Vacuolar protein sorting-associated protein 35 (hVPS35) for use in Western Blot, immunoprecipitation and immunofluorescence

F1000Res. 2023 Oct 16:12:452. doi: 10.12688/f1000research.133696.2. eCollection 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.

Keywords: Uniprot ID Q96QK1; VPS35; Vacuolar protein sorting-associated protein 35; Western Blot; antibody characterization; antibody validation; hVPS35; immunofluorescence; immunoprecipitation.

MeSH terms

  • Antibodies*
  • Blotting, Western
  • Fluorescent Antibody Technique
  • Immunoprecipitation
  • Protein Transport

Substances

  • Antibodies

Grants and funding

This work was supported by the Michael J. Fox Foundation for Parkinson’s Research (MJFF) (grant no. 18331). The Government of Canada through Genome Canada, Genome Quebec and Ontario Genomics (grant no. OGI-210). The Structural Genomics Consortium is a registered charity (no. 1097737) that receives funds from Bayer AG, Boehringer Ingelheim, Bristol Myers Squibb, Genentech, Genome Canada through Ontario Genomics Institute (grant no. OGI-196), the EU and EFPIA through the Innovative Medicines Initiative 2 Joint Undertaking (EUbOPEN grant no. 875510), Janssen, Merck KGaA (also known as EMD in Canada and the United States), Pfizer and Takeda. RA is supported by Mitacs fellowships.