| Solution Information | help | |
| Enzyme: | cAMP-dependent protein kinase catalytic subunit alpha | |
| inhibitor: | BDBM37895 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: Compounds identified from a previously described set of experiments entitled "Primary high-throughput assay for chemical inhibitors of protein kinase A (PKA) activity" (PubChem AID= 524) were selected for testing in this assay. Out of 94 compounds identified during the primary screen, all 94 compounds were assessed in dose response experiments in 10 point, 1:3 serial dilutions starting at a nominal test concentration of 60 micromolar.As with the primary screen, this assay is based on ability of PKA kinase to phosphorylate a Kemptide peptide sequence. PKA uses ATP as a donor of phosphate for the phosphorylation of the substrate, which leads to the depletion of ATP in the reaction mix. An assay kit (Kinase-Glo, Promega) was used to quantify enzyme activity. Residual amounts of ATP are measured by a secondary enzymatic reaction, during which luciferase utilizes the remaining ATP to produce luminescence. In this assay, the luminescent signal is directly proportional to | |
| If you find an error in this entry please send us an E-mail | ||