A high-throughput pipeline for validation of antibodies
Sikorski K, Mehta A, Inngjerdingen M, Thakor F, Kling S, Kalina T, Nyman TA, Stensland ME, Zhou W, de Souza GA, Holden L, Stuchly J, Templin M, Lund-Johansen F
Nat Methods. 2018 Nov;15(11):909-912. doi: 10.1038/s41592-018-0179-8. Epub 2018 Oct 30.
Western blotting (WB) is widely used to test antibody specificity, but the assay has low throughput and precision. Here we used preparative gel electrophoresis to develop a capture format for WB. Fractions with soluble, size-separated proteins facilitated parallel readout with antibody arrays, shotgun mass spectrometry (MS) and immunoprecipitation followed by MS (IP-MS). This pipeline provided the means for large-scale implementation of antibody validation concepts proposed by an international working group on antibody validation (IWGAV).