| Solution Information | help | |
| Enzyme: | Dual specificity protein phosphatase 3 | |
| inhibitor: | BDBM46223 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay materials: 1) VHR1 was obtained from the assay provider's laboratory. 2) Assay Buffer: 20mM Bis-Tris pH 6.0, 1mM DTT, 0.005% Tween-20 3) 3-O-Methylfluorescein phosphate (OMFP) cyclohexylammonium salt (Sigma-Aldrich, M2629) 4) VHR1 SC working solution: 4 nM VHR1 solution in the assay buffer 5) VHR1 DR working solution: 6 nM VHR1 solution in the assay buffer 6) OMFP SC working solution: 36 uM OMFP in the assay buffer 7) OMFP DR working solution: 25 uM OMFP in the assay buffer Procedure for single-concentration confirmation 1) Dispense 3 ul of working buffer in columns 1-2 of black 1536-well Corning plate (cat # 3724) 2) Dispense 3 uL of VHR1 SC working solution in columns 3-48. 3) Pin 20 nL of 2 mM compounds in DMSO in triplicate wells per compound to columns 3-46 or DMSO to columns 1-2 and 47-48. 4) Dispense 3 uL of OMFP SC working solution to wells in columns 1-48. 5) Briefly spin plate down 6) Incubate lidded plate for 1 hour at room temp. 7) Read plate on ViewLux | |
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