Commercial recombinant protein kinase fusion proteins were purchased from Medical Research Council Protein Phosphorylation and Ubiquitlyation Unit (PPUU) reagents (University of Dundee) and were purified from Sf21 cells or E. coli. Full details are provided in table S3. GST and His6-tagged kinases were purified using standard procedures, before storage in 1 mM DTT (except where indicated) at −80°C. All kinases were assayed using standard enzyme preparations (128). Bacterially expressed GST-MAPKAP-K2 and GST-MAPKAP-K3 were activated in vitro by incubation with catalytically active p38α, which was removed by repurification before assay. General biochemicals and all redox reagents, including GRX, were purchased from Sigma-Aldrich.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.