A high-throughput pipeline for validation of antibodies.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 30377371.
- Also identified by DOI 10.1038/s41592-018-0179-8.
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Abstract
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).
Medical subject headings
- Antibodies
- High-Throughput Screening Assays
- Neoplasm Proteins
- Neoplasms
- Proteomics