| Solution Information | help | |
| Enzyme: | Alkaline phosphatase, tissue-nonspecific isozyme | |
| inhibitor: | BDBM59303 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | 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 - 500 mM EDTA. 6) Positive Control (PC) solution - 10% DMSO. HTS protocol: 1) 2 uL of CDP-star working solution was added to all the well of Costar 1536-well white plate (cat #3725) using the Thermo WellMate dispenser 2) 40 nL of NC solution was added to columns 1-2 using V&P pin tool 3) 40 nL of PC solution was added to columns 3-4 using V&P pin tool 3) 40 nL of 2 mM compounds in 100% DMSO were dispensed in columns 5-48 4) 2 uL of TNAP working solution was added to the whole plate using the Thermo WellMate dispenser. 5) Final concentrations of the componen | |
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