| Solution Information | help | |
| Enzyme: | Tyrosine-protein phosphatase non-receptor type 22 | |
| inhibitor: | BDBM74492 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay materials: 1) LYP (LYPCAT) was produced at the Sanford-Burnham Medical Research Institute's protein production core facility 2) 14LckpCAP394 (pCAP) was purchased from Univ. of Utah peptide synthesis core 3) Assay Buffer = 50mM Bis Tris pH 6.0, 1mM DTT, 0.1% BSA and 0.005% Tween 20. The buffer was filtered prior to addition of enzyme or substrate. uHTS protocol: 1) Add 2uL of 50uM pCAP in assay buffer to all wells of the plate (final substrate concentration= 25uM) 2) Centrifuge plate 3) Using a 1536 pintool add 23nl 100% DMSO to columns 1-4 (control wells) 4) Using a 1536 pintool add 23nL of 2mM compound in DMSO for cols 5-48 (final compound concentration= 11.5uM) 5) Centrifuge plate 6) Add 2ul of assay buffer to wells in columns 1 and 2 (positive control) 7) Add 2uL of 8nM LYPCAT in assay buffer to wells in columns 3-48 (final enzyme concentration= 4nM) 8) Centrifuge plate 9) Incubate plates for 30 minutes at room temperature 10) Read plates on a Perkin Elmer Viewlux Dos | |
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