A high-throughput pipeline for validation of antibodies

用于抗体验证的高通量流程

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作者:Krzysztof Sikorski, Adi Mehta, Marit Inngjerdingen, Flourina Thakor, Simon Kling, Tomas Kalina, Tuula A Nyman, Maria Ekman Stensland, Wei Zhou, Gustavo A de Souza, Lars Holden, Jan Stuchly, Markus Templin, Fridtjof Lund-Johansen

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).

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