| Solution Information | help | |
| Enzyme: | Mevalonate kinase | |
| inhibitor: | BDBM38573 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Mevalonate Kinase Protocol for 384-well HTS Purified recombinant MK enzyme was provided by Dr. Thomas Leyh, Albert Einstein College of Medicine of Yeshiva University. Final Assay Conditions in 26 ul total volume: 50 mM HEPES buffer, pH 8.0 50 mM KCl 6 mM MgCl2 5 mM ATP 0.2 mM Mevalonate 2 mM Potassium Phosphoenolpyruvate (PEP) 0.4 mM NADH 3 units/ml rabbit muscle lactic dehydrogenase (LDH) 3 units/ml rabbit muscle pyruvate kinase 60 nM Mevalonate Kinase (MK) 4% DMSO Full reaction conditions are listed above. Background conditions are the same as above except that the substrate (mevalonate) is left out. The assay is run in 384 well clear non-binding plates. Fifteen ul substrate/background is added to plates using the BeckmanCoulter Biomek FX liquid handling workstation. Compounds are then added in 1 ul volumes using the FX. The reaction is initiated by the addition of 10 ul of MK using the Titertek Multidrop peristaltic pipetting platform. The test plate was immediately tran | |
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