| Assay Method Information | |
| | Activation Assay |
| Description: | In the 384-well plates 1.25 μl of test compound in assay buffer was mixed with 1.25 μl of AMPK and 1.25 μl of the peptide (final concentration of 1 μM) and 1.25 μl of ATP (final concentration of 30 μM), both dissolved in assay buffer. This step was followed by an incubation time of 60 min. Then 5 μl of ADP Glo Reagent was added. This was followed by 40 min of incubation. Then 10 μl of Kinase Detection Reagent was admixed. The plates were sealed and after an incubation period of 30 min, the luminescence signal was measured in an Envision reader. All incubation steps were accomplished at room temperature. Assay buffer: 20 mM HEPES pH 7.0, 0.025% BSA, 15 mM MgCl2, 0.01% Brij. |
| Affinity data for this assay | |
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