Investigating the applicability of antibodies generated within the human protein atlas as capture agents in immunoenrichment coupled to mass spectrometry.
作者： Tove Boström , Henrik J. Johansson , Janne Lehtiö , Mathias Uhlén , Sophia Hober
DOI: 10.1021/PR500691A
关键词:
摘要: For identification and characterization of proteins in complex samples, immunoenrichment coupled to mass spectrometry is a good alternative due the sensitivity affinity enrichment specificity analysis. Antibodies are commonly used agents; however, for high-throughput analysis, antibody availability usually bottleneck. Here we present protocol setup, where all steps from bead coupling sample preparation performed parallel 96-well format. generated within Human Protein Atlas project were tested applicability as capture agents. The antibodies covalently attached protein A beads, making it possible reuse beads at least three times without destroying binding efficiency. Target captured U251 MG cell lysate, eluted, digested, analyzed using spectrometry. Of 30 investigated...
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