Solution Informationhelp
Enzyme: RAC-alpha serine/threonine-protein kinase
inhibitor: BDBM32361
substrate: n/a
 
Solution Type: Aqueous
pH at Preparation: n/a
Temp. Prep.: n/a
Comments: Protocol AKT counterscreening assay protocol for identified PKD inhibitors (384-well HTS) Reagents 1X kinase reaction buffer 10 mM Tris-HCl, pH 7.2 10 mM MgCl2 0.05% NaN3 0.1% Bovine serum albumin 1 mM DTT Substrate/ATP (3X concentration) 900 nM substrate peptide 30 uM ATP in 1X kinase reaction buffer Max control (3X concentration) 3% DMSO in 1X kinase reaction buffer Min control (3X concentration) 300 uM H-89 in 1X kinase reaction buffer IC50 control (3X concentration) 15 uM H-89 in 1X kinase reaction buffer Compounds (3X concentration) 2 uL of 1 mM compounds solubilized in 1X kinase reaction buffer starting at 300 uM (3X concentration) and subjected to a 2-fold serial dilution Enzyme (3X concentration) 0.15 mUnits/uL in 1X kinase reaction buffer Binding reagent 1:400 dilution binding reagent in 75% 1X binding buffer A and 25% 1X binding buffer B Assay Procedure (1)Prepare reagents as described above (2)Add 2 uL of a substrate peptide and ATP mixture to appropriate wells.
 
 
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