A guide to selecting high-performing antibodies for Protein-glutamine gamma-glutamyltransferase 2 (TGM2) for use in western blot, immunoprecipitation and immunofluorescence

F1000Res. 2024 Jul 30:13:481. doi: 10.12688/f1000research.150684.2. eCollection 2024.

Abstract

Protein-glutamine gamma-glutamyltransferase 2 (TGM2) is a Ca 2+ dependent enzyme that catalyzes transglutaminase cross-linking modifications. TGM2 is involved in various diseases, either in a protective or contributory manner, making it a crucial protein to study and determine its therapeutic potential. Identifying high-performing TGM2 antibodies would facilitate these investigations. Here we have characterized seventeen TGM2 commercial antibodies for western blot and sixteen for immunoprecipitation, and immunofluorescence. The implemented standardized experimental protocol is based on comparing read-outs in knockout cell lines against their isogenic parental controls. This study is part of a larger, collaborative initiative seeking to address antibody reproducibility issues by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While the use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.

Keywords: Protein-glutamine gamma-glutamyltransferase 2; TGM; TGM2; Uniprot ID P21980; antibody characterization; antibody validation; immunofluorescence; immunoprecipitation; western blot.

MeSH terms

  • Antibodies* / immunology
  • Blotting, Western*
  • Fluorescent Antibody Technique* / methods
  • GTP-Binding Proteins / immunology
  • Humans
  • Immunoprecipitation* / methods
  • Protein Glutamine gamma Glutamyltransferase 2*
  • Transglutaminases* / immunology

Substances

  • Transglutaminases
  • Protein Glutamine gamma Glutamyltransferase 2
  • Antibodies
  • GTP-Binding Proteins

Grants and funding

This work was supported by the Michael J. Fox Foundation for Parkinson’s Research (MJFF) (grant no. 18331). This work was also supported by a grant from the Canadian Institutes of Health Research Foundation (grant no. FDN154305), the Government of Canada through Genome Canada, Genome Quebec and Ontario Genomics (grant no. OGI-210). RA is supported by Mitacs fellowship.