| Assay Method Information | |
| | CtBP2 ITC |
| Description: | Purified CtBP2 was dialyzed overnight against 500 mL of 50 mM glycylglycine at pH 8.5, 300 mM NaCl, 1 mM EDTA, and 2 mM TCEP, with or without 1.5 mM NAD+.Concentrated ligands were diluted to working concentrations in dialysate. Binding was measured by titrating in 1.5 mM HIPP. Five mM MTOB was utilized for competitive displacement assays. MTOB was incubated with the protein sample and HIPP for 10 min prior to the displacement assays. 1.5 mM HIPP was titrated to displace MTOB from the CtBP2 active site. CtBP2 concentration was 100−120 μM forall experiments. Each measurement consisted of 20−25 1.0 μL injections of HIPP into the CtBP2 solutions. |
| Affinity data for this assay | |
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