| Solution Information | help | |
| Enzyme: | Alkaline phosphatase, germ cell type | |
| inhibitor: | BDBM41892 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Materials: 1)Human IAP protein was provided by Dr. Jose Luis Millan (Sanford-Burnham Medical Research Institute, San Diego, CA). The CDP-star was obtained from Applied Biosystems. 2)Assay Buffer: 250 mM DEA, pH 9.8, 2.5 mM MgCl2, and 0.05 mM ZnCl2. 3)IAP working solution contained a 1/80 dilution in assay buffer. Solution was prepared 1h prior to use and incubated at room temperature. 4)CDP-star working solution contained 442.5 uM CDP-star in MQ water. 5)L-phenylalanine working solution - 100 mM in 10% DMSO. IAP dose-response confirmation screening 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 white small-volume plates (784075). Each curve was performed in duplicate. 2)Columns 1-2 and 23-24 contained 4 uL of L-phenylalanine workin | |
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