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Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods

Article DOI: 10.3791/59795-v • 07:49 min • July 17th, 2019
July 17th, 2019 •
1, 1, 1, 1, 1, 1,2
1The Laboratory of Cancer Signaling, Division of Cellular and Molecular Research, National Cancer Centre Singapore, 2The Cancer and Stem Cell Program, Duke-NUS Medical School

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In this article, we presented a set of practical and feasible methods for characterizing disease-related mutants of RAF family kinases, which include in vitro kinase assay, RAF co-activation assay, and complementary split luciferase assay.

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Disease-related Mutants RAF Family Kinases Practical Methods Feasible Methods Characterization Catalytic Activity Flag Tech Mutations PCR Vector Glutathione As Transferase 293T Cell Cultures Lysis Buffer Protease Inhibitors Phosphatase Inhibitors Centrifugation Cell Debris Micro Centrifuge Tubes Downstream Analysis Extracellular Signal Regulated Kinase
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