| Assay Method Information | |
| | In vitro Enzyme Assay |
| Description: | Reaction Biology Corporation Kinase HotSpotSM service: In a final reactionvolume of 25 μL, kinase (5-10 mU) is incubated with 25 mM Tris pH 7.5, 0.02 mM ethylene glycol tetracetic acid (EGTA), 0.66 mg/mL myelin basic protein, 10 μM magnesium acetate, and [γ33P-ATP] (specific activity approx. 500 c.p.m./pmol,concentration as required). The reaction is initiated by the addition of the Mg-ATP mix. After incubation for 40 min at room temperature, the reaction is stopped by the addition of 5 μL of a 3% phosphoric acid solution. About 10 μL ofthe reaction solution is then spotted onto a P30 filtermat and washed three times for 5 min in 75 mM phosphoric acid and once in methanol prior to drying and scintillation counting. |
| Affinity data for this assay | |
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