| Assay Method Information | |
| | In Vitro Inhibition Assay |
| Description: | The kinase activity of CaMKII beta is evaluated by means of a radioactive test on the recombinant CaMKII beta enzyme. The amount of ATP-gamma 33P incorporated into the specific substrate Autocamtide-2 during its phosphorylation by CaMKII beta is measured. The effect of the products is quantified by the concentration of product that inhibits the total activity of CaMKII beta by 50% (50% inhibitory concentration=IC50). For the determination of the IC50 values, the product is diluted in 100% of DMSO to obtain a 10 mM stock solution. The concentration range tested during the radioactive test ranges from 3 to 10 000 nM with a final concentration in the test of 1% DMSO. This concentration range may, for the most powerful compounds, be extended to 0.1 nM. On the day of the operation, 5 ul of the compounds are deposited in each well of a 96-well plate at 10 times the concentration of that to be tested. Each concentration is tested in duplicate on the same plate. |
| Affinity data for this assay | |
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