| Assay Method Information | |
| | Kinase Inhibition Assay |
| Description: | Method for PIM2 Kinase Inhibition Assay: Dowex Technique i. Kinase Buffer (KB)The buffer for PIM2 assay was composed of HEPES 50 mM, at pH 7.5, with 1 mM MgCl2, 1 mM DTT, 3 uM Na3VO4, and 0.2 mg/mL BSA.Full-length human PIM2 was expressed and purified as described in Fedorov O, et al., PNAS 2007 104, 51, 20523-28 ii. Assay Conditions (Final Concentrations)Enzyme concentration=1.5 nMAktide substrate (Chemical Abstract Service Registry Number 324029-01-8)=5 uM ATP=4 uM. 33P gamma ATP=1 nM iii. Robotized Dowex AssaySee above: same procedure as described for PIM1. |
| Affinity data for this assay | |
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