Assay Method Information

Assay Name:  TrkA Enzyme Activity Test
Description:  In a 10 μL reaction system with a buffer of 20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.01% Brij35, 0.1 mM sodium vanadate, 0.02 g/mL BSA, 2 mM DTT, 1% DMSO medium, containing 15 nM TrkA kinase, 0.3 μM biotin-TK peptide (biotin-labeled tyrosine kinase substrate polypeptide), 100 μM ATP were incubated at 23° C. for 120 minutes. The reaction was spotted on P81 ion exchange paper (Whatman #3698-915), the filter was washed thoroughly with 0.75% phosphoric acid, and the radiophosphorylated substrate remaining on the filter was measured. Kinase activity data were expressed as a percentage of kinase activity in the test sample compared to the vehicle (DMSO) reaction. IC50 and curve fitting can be obtained by Graphpad software Prism4.
Affinity data for this assay
 

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