| Solution Information | help | |
| Enzyme: | Glyceraldehyde-3-phosphate dehydrogenase | |
| inhibitor: | BDBM42075 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | GAPDH assay materials: 1)rabbit GAPDH, NAD+ and glyceraldehydes-3-phosphate were purchased from Sigma-Aldrich 2)Assay Buffer: 62.5 mM glycine, 62.5 mM phosphate, pH 8.9, containing 2.5 mM EDTA, 2.5 mM NAD+, and 2.5 mM DTT. 3)GAPDH working solution: 12 nM GAPDH in the assay buffer 4)G3P working solution: 0.735 mM glyceraldehyde-3-phosphate solution in water. GAPDH dose response assay protocol: 1) 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 water to 10% final DMSO concentration. 4 uL compounds in 10% DMSO were transferred into columns 3-22 of Greiner 384-well clear plates (781101). Columns 1-2 and 23-24 were added with 4 uL of 0.15% H2O2 and 10% DMSO. 2) 8 uL of GAPDH working solution was added to columns 1-24 using WellMate bulk dispenser (Matrix). 3) 8 uL of G3P working solution was added to columns 1-24 using WellMate bulk dispenser (Matrix). 4) 8 uL of G3P w | |
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