| Solution Information | help | |
| Enzyme: | Alkaline phosphatase, tissue-nonspecific isozyme | |
| inhibitor: | BDBM44387 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | TNAP assay protocol. 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 Applied Biosystems. 2) Assay Buffer: 250 mM DEA, pH 9.8, 2.5 mM MgCl2, and 0.05 mM ZnCl2. 3) TNAP working solution contained a 1/800 dilution in assay buffer. Solution was prepared fresh prior to use. 4) CDP-star working solution contained 125 uM CDP-star in MQ water. 5) Levamisole working solution - 5 mM in 10% DMSO. TNAP HTS protocol: 1) 4 uL of 100 uM compounds in 10% DMSO were dispensed in columns 3-24 of Greiner 384-well white small volume plates (784075). 2) Using the Thermo wellmate dispenser 4 uL the following solutions were added: a. 10% DMSO - column 1 (negative control). b. Levamisole working solution - columns 2 (positive control). 3) 8 uL of TNAP working solution was added to the whole plate using WellMate bulk dispenser (Matrix). 4) 8 uL of CDP-star working solution was added to | |
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