| Solution Information | help | |
| Enzyme: | Alkaline phosphatase, tissue-nonspecific isozyme | |
| inhibitor: | BDBM43792 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | TNAP assay materials: 1) TNAP 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: 100 mM CAPS, pH 9.8, 2 mM MgCl2, and 0.04 mM ZnCl2. 3) TNAP working solution contained a 1/400 dilution in assay buffer. 4) CDP-star working solution contained 200 uM CDP-star in MQ water. 5) Negative Control (NC) solution - 5 mM levamisole. 6) Positive Control (PC) solution - 10% DMSO. TNAP 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 NC and PC solutions, respectively. 3 | |
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