| Solution Information | help | |
| Enzyme: | Tyrosine-protein phosphatase YopH | |
| inhibitor: | BDBM42131 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Protocol YopH Phosphatase Activity Protocol for 384-well HTS Solutions 3X Assay buffer 30 mM Magnesium Chloride 150 mM HEPES 3 mM Dithiothreitol 0.3% BSA (pH ~7.0) (3X) MAX control 3% DMSO in water (3X) MIN control 600 mM sodium hydroxide in water (3X) IC50 control 75 mM sodium hydroxide in water (3X) Substrate 75 uM OMFP in water (3X) Stop Solution 600 mM sodium hydroxide in water (3X) Test compounds 150 uM compounds diluted in water (starting concentration for two-fold serial dilution) (3X) YopH enzyme 200 ng/mL in 3X assay buffer Assay procedure 1.Prepare the following reagents in the appropriate diluent. 2.Transfer 5 uL of the diluted compounds (30 uM) and plate controls to wells of a Greiner 384-well, low-volume, assay plate. 3.Add 5 uL enzyme (200 ng/mL) and 5 uL substrate (75 uM) using a Flexdrop bulk dispenser. 4.Centrifuge plates at 500 g X 1 minute. 5.Incubate plates for 1 hour at room temperature. 6.Stop the assay reaction by adding 5 uL of stop solution to e | |
| If you find an error in this entry please send us an E-mail | ||