A high-throughput pipeline for validation of antibodies
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
30377371
DOI
10.1038/s41592-018-0179-8
PII: 10.1038/s41592-018-0179-8
Knihovny.cz E-resources
- MeSH
- Mass Spectrometry MeSH
- Immunoprecipitation MeSH
- Humans MeSH
- Tumor Cells, Cultured MeSH
- Neoplasm Proteins immunology metabolism MeSH
- Neoplasms immunology metabolism MeSH
- Proteomics methods MeSH
- Antibodies immunology MeSH
- High-Throughput Screening Assays methods standards MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Neoplasm Proteins MeSH
- Antibodies MeSH
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).
Department of Immunology University of Oslo and Oslo University Hospital Oslo Norway
Department of Pathology Oslo University Hospital Oslo Norway
K G Jebsen Center for Cancer Immunotherapy University of Oslo Oslo Norway
K G Jebsen Inflammation Research Centre University of Oslo Oslo Norway
NMI Natural and Medical Sciences Institute University of Tübingen Reutlingen Germany
Norwegian Computing Center Oslo Norway
SeekQuence Mountain View CA USA
The Brain Institute Universidade Federal do Rio Grande do Norte Natal Brazil
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