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Table 2 IWGAV strategy for antibody validation

From: Minimum Information and Quality Standards for Conducting, Reporting, and Organizing In Vitro Research

Strategy Model Number of antibodies Analysis
Genetic Knockout or knockdown cells/tissues Antibody of interest Antibody-based method of choice
Orthogonal Several samples Antibody of interest Correlation between antibody-based and antibody-independent assays
Independent antibodies Lysate or tissue with target protein Several independent antibodies with different epitopes Specificity analysis through comparative and quantitative analysis
Tagged proteins Lysate or tissue containing tagged and native protein Anti-tag antibody compared with antibody of interest Correlating the signal from the tagged and non-tagged proteins
Immunocapture followed by MS Lysate containing protein of interest Antibody of interest Target immunocapture and mass spectrometry of target and potential binding partners