| Solution Information | help | |
| Enzyme: | Tyrosine-protein phosphatase non-receptor type 7 | |
| inhibitor: | BDBM75372 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | HePTP assay materials: 1)HePTP protein was provided by Dr. Mustelin (Sanford-Burnham Medical Research Institute, San Diego, CA). 2)Assay Buffer: 50 mM Bis-Tris, pH 6.0, 375 mM NaCl, 2.5 mM DTT, 0.0125% Tween 20. 3)HePTP working solution contained 6.875 nM HePTP in assay buffer. Solution was prepared fresh prior to use. 4)The OMFP 20 mM stock solution was prepared by dissolving 10.5 mg OMFP in 1 mL DMSO and sonicating the solution for 1 min. Right before use, the stock solution was diluted to a working solution of 750 uM concentration. 5)Vanadate working solution - 45 mM Na3VO4 in 10% DMSO HePTP dose-response assay 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). Columns 1-2 and 23-24 contained 4 uL | |
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