Survey of commercial antibodies targeting Y chromosome-encoded genes

Abstract Commercial antibodies are both a mainstay of biomedical research and subject to increasingly widespread scrutiny for their specificity. We evaluated the specificity of commercially available research grade antibodies by investigating those whose antigen targets are encoded by the Y chromosome. We assessed vendor-provided validation data such as immunoblotting and identified 65 commercial research grade antibodies that exhibited positive reactivity in female-derived tissues and/or cell lines. A thorough analysis of 30 commercial antibodies marketed as targeting the Y chromosome-encoded protein DDX3Y found that 16 (53%) provided no marketing data on specificity, 9 (30%) provided positive data in female or likely female materials, 4 (13%) provided only positive data in male materials, and 1 (3%) provided positive data in male materials and negative data in female materials. Together, these findings suggest that many commercial antibodies may be unsuitable to distinguish between related X and Y chromosome-encoded gene pairs and should instill caution in researchers using these tools to investigate sex chromosome-encoded proteins.