Vitenskapelig artikkel

A high-throughput pipeline for validation of antibodies

Sikorski, Krzysztof; Mehta, Adi; Inngjerdingen, Marit; Thakor, Flourina Kumar; Kling, Simon; Kalina, Tomas; Nyman, Tuula Anneli; Stensland, Maria; Zhou, Wei; De Souza, Gustavo Antonio; Holden, Lars; Stuchly, Jan; Templin, Markus; Lund-Johansen, Fridtjof

Publikasjonsdetaljer

Tidsskrift: Nature Methods, vol. 15, p. 909–912, 2018

Utgave: 11

Internasjonale standardnumre:
Trykt: 1548-7091
Elektronisk: 1548-7105

Lenker:
ARKIV: http://hdl.handle.net/10852/67592
DOI: doi.org/10.1038/s41592-018-0179-8

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).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).