| Solution Information | help | |
| Enzyme: | Acetyl-CoA acetyltransferase/HMG-CoA reductase | |
| inhibitor: | BDBM42077 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | HMG-CoA Reductase Protocol for 384-well HTS Purified recombinant Enterococcus faecalis HMG-CoA reductase was provided by Drs. Cynthia Stauffacher and Victor Rodwell at Purdue University. Compound Dosing/Plating: For the dose response screen, a 1:2 serial dilution in DMSO was made from the source plate, and 1 ul of compound was dispensed into assay plates, resulting in a final concentration range of 100-0.2 uM. Assay Setup: Fifteen uL of HMG-CoA reductase reagent mix which included coenzyme A, NADP, and mevalonate in assay buffer was added to each well of the previously compound dosed 384-well plates. The reaction was initiated with the addition of 10 uL of HMG-CoA reductase diluted in assay buffer. The final concentrations in the reaction were 2 mM coenzyme A, 4 mM NADP, 4 mM mevalonate, and 15 ug/ml HMG-CoA reductase diluted in assay buffer (100 mM Tris-HCl (pH 8), 100 mM KCl, 2% DMSO and 0.01% Tween 20). The test plate was immediately transferred to a Perkin Elmer Envision micro | |
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