| Assay Method Information | |
| | Enzymatic Inhibition of TTBK1 and TTBK2 |
| Description: | A buffer solution containing the recombinant human enzyme TTBK1 (1-1321) or TTBK2(1-450) (5-20 mU) and 50 mM Tris at pH 7.5, 0.1 mM EGTA, 0.1% p-mercaptoethanol, 1 mg/ml BSA, 10 mM DTT, is tested against the peptide RRKDLHDDEEDEAMSITA (SEQ ID NO: 1) in a final volume of 25.5 μl. The final reaction mixture contains the peptide at a concentration of 0.3 mM, 10 mM magnesium acetate and 0.005 mM [33P-γ-ATP](50-1000 cpm/pmol). The reaction mixture is incubated for 30 minutes at room temperature and the reaction is stopped after the addition of 5 μl of orthophosphoric acid. The mixture is placed on P81 Unifilter plates and washed with 50 mM orthophosphoric acid for data reading. |
| Affinity data for this assay | |
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