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Antibody validation for Western blot: By the user, for the user

The Journal of biological chemistry, 2020-01, Vol.295 (4), p.926-939 [Peer Reviewed Journal]

2020 Pillai-Kastoori et al. 2020 Pillai-Kastoori et al. ;ISSN: 0021-9258 ;EISSN: 1083-351X ;DOI: 10.1074/jbc.RA119.010472 ;PMID: 31819006

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  • Title:
    Antibody validation for Western blot: By the user, for the user
  • Author: Pillai-Kastoori, Lakshmi ; Heaton, Sam ; Shiflett, Steve D. ; Roberts, Annabelle C. ; Solache, Alejandra ; Schutz-Geschwender, Amy R.
  • Subjects: Methods and Resources
  • Is Part Of: The Journal of biological chemistry, 2020-01, Vol.295 (4), p.926-939
  • Description: Well-characterized antibody reagents play a key role in the reproducibility of research findings, and inconsistent antibody performance leads to variability in Western blotting and other immunoassays. The current lack of clear, accepted standards for antibody validation and reporting of experimental details contributes to this problem. Because the performance of primary antibodies is strongly influenced by assay context, recommendations for validation and usage are unique to each type of immunoassay. Practical strategies are proposed for the validation of primary antibody specificity, selectivity, and reproducibility using Western blot analysis. The antibody should produce reproducible results within and between Western blotting experiments and the observed effect confirmed with a complementary or orthogonal method. Routine implementation of standardized antibody validation and reporting in immunoassays such as Western blotting may promote improved reproducibility across the global life sciences community.
  • Publisher: 11200 Rockville Pike, Suite 302, Rockville, MD 20852-3110, U.S.A: American Society for Biochemistry and Molecular Biology
  • Language: English
  • Identifier: ISSN: 0021-9258
    EISSN: 1083-351X
    DOI: 10.1074/jbc.RA119.010472
    PMID: 31819006
  • Source: PubMed Central
    Alma/SFX Local Collection
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