| Solution Information | help | |
| Enzyme: | Alkaline phosphatase, germ cell type | |
| inhibitor: | BDBM39653 | |
| 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) Negative Control (NC) solution - 5 mM TCEP in 10% DMSO. 6) Positive Control (PC) solution - 10% DMSO. PLAP HTS protocol: 1) 4 uL of NC solution was added to column 1 using the Thermo wellmate dispenser 2) 4 uL of PC solution was added to column 2 using the Thermo wellmate dispenser 3) 4 uL of 100 uM compounds in 10% DMSO were dispensed in columns 3-24 of Greiner 384-well white small volume plates (784075) using BioMek FX. 4) 8 uL of PLAP working solution was added to the whole plate using WellMate bulk dispenser (Matrix). 5) 8 uL of CDP-star worki | |
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