The target of phosphorylation of CDK5 is MAP1B, so an specific antibody for the phosphorylated MAP1B should constitute a very good assay for measuring the activity of CDK5, in the page is the announcement of the launch of this type of antibodies, it´s from 2010 so I think more supliers should have developed this useful protein.
http://www.novusbio.com/about/press-release/novus-biologicals-announces-new-phospho-map1b-antibody
You can measure kinase activity on a peptide microarray. See www.pamgene.com for more information
Thanks Dr. Hilhorst, however I could not find information how to measure CDK5 kinase activity in the cell lysate. I just checked the suggested website.
Following Dr. Rodriguez's suggestion, I contacted novusbio. However, novusbio confirmed that they do not have cdk5 assay related product. My understanding from literature is that histone is the main target, and researchers have used this to assay cdk5 activity. Thanks Dr. Rodriguez.
Hello Navin,
when you go to https://www.pamgene.com/en/publications.htm you find quite a number of publications.This one illustrates the measurement of activity of a recombinant kinase. CDK5 activity can probalby also be measured. The second one illustrates measuring kinase activity in tissue.
Peptide microarrays for detailed, high-throughput substrate identification, kinetic characterization, and inhibition studies on protein kinase A.
Hilhorst R, Houkes L, van den Berg A, Ruijtenbeek R. (2009) Anal Biochem. 2009 Apr 15;387(2):150-61. Epub 2009 Jan 21.
Kinome profiling in pediatric brain tumors as a new approach for target discovery.
Sikkema AH, Diks SH, den Dunnen WF, ter Elst A, Scherpen FJ, Hoving EW, Ruijtenbeek R, Boender PJ, de Wijn R, Kamps WA, Peppelenbosch MP, de Bont ES. (2009) Cancer Res. 2009 Jul 15;69(14):5987-95. Epub 2009 Jun 30.
Try Kinase-Glo ® Luminescent Kinase Assay ,
Best,
Thiru
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