A high-throughput pipeline for validation of antibodies

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).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / immunology*
  • High-Throughput Screening Assays / methods*
  • High-Throughput Screening Assays / standards*
  • Humans
  • Immunoprecipitation
  • Mass Spectrometry
  • Neoplasm Proteins / immunology*
  • Neoplasm Proteins / metabolism
  • Neoplasms / immunology
  • Neoplasms / metabolism*
  • Proteomics / methods*
  • Tumor Cells, Cultured


  • Antibodies
  • Neoplasm Proteins