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  • Identifying Kinase Substrates via a Heavy ATP Kinase Assay and Quantitative Mass Spectrometry.

Identifying Kinase Substrates via a Heavy ATP Kinase Assay and Quantitative Mass Spectrometry.

Scientific reports (2016-06-28)
André C Müller, Roberto Giambruno, Juliane Weißer, Peter Májek, Alexandre Hofer, Johannes W Bigenzahn, Giulio Superti-Furga, Henning J Jessen, Keiryn L Bennett
摘要

Mass spectrometry-based in vitro kinase screens play an essential role in the discovery of kinase substrates, however, many suffer from biological and technical noise or necessitate genetically-altered enzyme-cofactor systems. We describe a method that combines stable γ-[(18)O2]-ATP with classical in vitro kinase assays within a contemporary quantitative proteomic workflow. Our approach improved detection of known substrates of the non-receptor tyrosine kinase ABL1; and identified potential, new in vitro substrates.

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Sigma-Aldrich
单克隆抗-HA 小鼠抗, clone HA-7, ascites fluid
Sigma-Aldrich
Anti-Phosphotyrosine Antibody, recombinant clone 4G10®, clone 4G10®, Upstate®, from mouse