| Solution Information | help | |
| Enzyme: | Phosphomannomutase 2 | |
| inhibitor: | BDBM34233 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | PMM assay materials: 1) Human PMM2 protein was provided by Dr. Hudson Freeze (Sanford-Burnham Medical Research Institute, San Diego, CA). 2) Substrate working solution: 50 mM HEPES, pH 7.4, 0.166 mM Mannose-1-phosphate, 0.411 U/ml Diaphorase, 0.056 mM Resazurin. 3) Enzyme working solution: 50 mM HEPES, pH 7.4, 1.11 mM NADP+, 10.0 mM MgCl2, 0.011% Tween 20, 5.0 ug/ml phosphoglucose isomerase, 3.33 ug/ml PMI, 2.0 ug/ml G6PDH, and 1.67 ug/ml PMM2, and 0.11 mM alpha-D-Glucose 1,6-bisphosphate. PMM2 Dose-response confirmation protocol: 1) 9 uL of Substrate working solution was added to columns 3-24 of a Greiner 384-well black plate (cat # 784076) using a Multidrop Combi liquid dispenser (Thermo) 2) 9 ul of Substrate working solution without mannose-1-p was added to columns 1 and 2 (positive control) 3) Dose-response curves contained 10 concentrations of compounds obtained using 2-fold serial dilution. Compounds were serially diluted in 100% DMSO, and then diluted with 50 mM HEPES, pH | |
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