| Solution Information | help | |
| Enzyme: | Tyrosine-protein phosphatase non-receptor type 22 | |
| inhibitor: | BDBM32025 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Material - Recombinant LYP protein was provided by Dr. Mustelin (Burnham Institute for Medical Research, La Jolla, CA) - Bis-Tris (Sigma-Aldrich, B4429-500G) - NaCl (Sigma-Aldrich, S-7653) - DTT (Sigma-Aldrich, D-9779) - PEG (polyethylene glycol) 8000 (Sigma-Aldrich, P2139) - DiFMUP (Invitrogen, D22065) - Na Orthovanadate (Calbiochem, 567540) Buffers (Batch size of 205 plates) 1. Assay Buffer: 0.15 M Bis-Tris, pH 6.0, bring to ionic strength of 150 mM by addition of NaCl. Filter sterilize (0.22 micron, Corning 430186) 2. Enzyme Buffer: Assay buffer, 1% PEG, 5 mM DTT. DTT is added immediately before use. 3. LYP working solution: 7.5 nM LYP in enzyme buffer. Solution was prepared fresh prior to use. 4. DIFMUP working solution: 150 uM DIFMUP in assay buffer. Solution was prepared fresh prior to use and protected from light. 5. Orthovanadate working solution: 8 mM Na orthovanadate in assay buffer. Procedure NOTE: Assay must be performed in the dark because DiFMUP is l | |
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