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

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  • Additional Information
    • Corporate Authors:
      Neuro/SGC/EDDU collaborative group; ABIF consortium
    • Source:
      Publisher: F1000 Research Ltd Country of Publication: England NLM ID: 101594320 Publication Model: eCollection Cited Medium: Internet ISSN: 2046-1402 (Electronic) Linking ISSN: 20461402 NLM ISO Abbreviation: F1000Res Subsets: MEDLINE
    • Publication Information:
      Original Publication: London : F1000 Research Ltd
    • Subject Terms:
      Transglutaminases*/immunology ; Blotting, Western* ; Fluorescent Antibody Technique*/methods ; Protein Glutamine gamma Glutamyltransferase 2* ; Immunoprecipitation*/methods ; Antibodies*/immunology; Humans ; GTP-Binding Proteins/immunology
    • 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.
      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: - ABCD antibodies - Abcam - Aviva Systems Biology -Bio Techne -Cell Signalling Technology -Developmental Studies Hybridoma Bank -GeneTex – Horizon Discovery – Proteintech – Synaptic Systems –Thermo Fisher Scientific.
      (Copyright: © 2024 Ayoubi R et al.)
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    • Contributed Indexing:
      Keywords: Protein-glutamine gamma-glutamyltransferase 2; TGM; TGM2; Uniprot ID P21980; antibody characterization; antibody validation; immunofluorescence; immunoprecipitation; western blot
    • Accession Number:
      EC 2.3.2.13 (Transglutaminases)
      EC 2.3.2.13 (Protein Glutamine gamma Glutamyltransferase 2)
      0 (Antibodies)
      EC 3.6.1.- (GTP-Binding Proteins)
    • Publication Date:
      Date Created: 20240902 Date Completed: 20240902 Latest Revision: 20240903
    • Publication Date:
      20240903
    • Accession Number:
      PMC11362715
    • Accession Number:
      10.12688/f1000research.150684.2
    • Accession Number:
      39220380