Solution Informationhelp
Enzyme: Serine/threonine-protein kinase 33
inhibitor: BDBM67602
substrate: n/a
 
Solution Type: Aqueous
pH at Preparation: n/a
Temp. Prep.: n/a
Comments: Assay Buffer (AB) 10 mM MOPS pH 7 300 uM EDTA 0.5 % (V/V) Glycerol 0.001% (V/V) Brij-35 0.1 mg/mL BSA 0.01% (V/V) beta mercapto ethanol ATP/Mg 250 uM ATP 50 mM MgAc2 in AB STK33/MBP (E) 50 nM STK33 0.5 mg/mL Myelin Basic Protein in AB 1536 well plate assay Add 2.0 uL AB to last two columns of 1536 well white Aurora high-base assay plate (containing 2.5 nL 10 mM compound or DMSO) using combi-nL dispenser (Thermo). Add 2.0 uL E to all but last two columns of 1536 ARP (containing 2.5 nL 10 mM compound/well). Incubate 15 minutes at room temperature. Add 0.5 uL ATP/Mg / well to full plate. Incubate 60 minutes. Add 2.5 uL / well ADP Glo Reagent I (Promega). Incubate 40 minutes. Add 5 uL / well ADP Glo Reagent II (Promega). Incubate 30 minutes. Read luminescence 15 sec integration time on ViewLux plate reader (Perkin Elmer). 384 well plate assay Add 20 uL AB to last column of 384 well white Aurora assay plate using combi dispenser (Thermo). Add 20 uL E to all but las
 
 
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