| Solution Information | help | |
| Enzyme: | Alkaline phosphatase, germ cell type | |
| inhibitor: | BDBM46053 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | PLAP assay materials: 1) PLAP protein was provided by Dr. Jose Luis Millan (Sanford-Burnham Medical Research Institute, San Diego, CA). The CDP-star was obtained from New England Biolabs. 2) Assay Buffer: 250 mM DEA, pH 9.8, 2.5 mM MgCl2, and 0.05 mM ZnCl. 3) PLAP working solution contained a 1/6400 dilution in assay buffer. Solution was prepared fresh prior to use. 4) CDP-star working solution contained 212.5 uM CDP-star in MQ water. 5) TCEP working solution - 5 mM in 10% DMSO. PLAP 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). Columns 1-2 and 23-24 contained 4 uL of TCEP working solution and 10% DMSO, respectively. 2) 8 uL of PLAP working solution was adde | |
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