In vitro JAK kinase activity and inhibition assays.
Author(s)
Babon, JJ; Murphy, JM;
Journal Title
Methods in Molecular Biology
Publication Type
Journal Article
Abstract
The discovery that a range of myeloproliferative diseases and leukemias are associated with Janus Kinase (JAK) mutations has highlighted the importance of JAK/STAT signalling in disease and sparked a renewed interest in developing JAK inhibitors. In vitro kinase assays are the most direct and quantitative method to assess mutant forms of JAK for altered enzymatic properties as well as verifying and quantifying the affinity and efficacy of potential inhibitors. Here, we describe protocols for heterologous expression and purification of JAK kinases from insect cells, assays to determine the activity of these purified kinases, and finally inhibition assays to determine the effectiveness of potential inhibitors.
Publisher
Springer
Keywords
JAK; Kinase ; Enzyme ; Inhibition assay ; Activation ; Enzyme kinetics
Research Division(s)
Structural Biology; Cancer And Haematology
PubMed ID
23296720
Terms of Use/Rights Notice
© Springer, Part of Springer Science+Business Media


Creation Date: 2013-01-01 12:00:00
Last Modified: 2018-01-16 10:06:24
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