| Solution Information | help | |
| Enzyme: | Mothers against decapentaplegic homolog 3 | |
| inhibitor: | BDBM40315 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Compounds identified as active from the primary screen, AID 630 - HTS of SMAD transcription factor inhibitors, were selected for testing in this assay. Each compound was tested in 6 point, 1:2 serial dilutions starting at a nominal test concentration of 125 micromolar. Each concentration was tested in triplicate. In some cases, compounds were retested in 6 point, 1:2 serial dilutions starting at lower concentrations, to obtain the final IC50. These cases are noted in the PUBCHEM_ASSAYDATA_COMMENT field. Reagents: 1. Assay buffer: K-Hepes, 50 mM, pH 7.0; KCl, 100 mM; BSA, 1mg/ml; DTT, 1mM; Tween 20, 0.2%. 2. Phosphorylated, 6xHis-tagged Smad3 protein: purified from an insect cell expression system as previously described (Cui, Q., et al, 2005; Oncogene, 24: 3864-74) 3. Dy647-hFoxH1 peptide: synthesized at the University of Wisconsin-Madison Biotechnology Center and dissolved in DMF 4. Europium Chelate Anti-6xHis-Antibody: LANCE Eu-W1024 from Perkin Elmer Life Sciences 5. Posit | |
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