| Solution Information | help | |
| Enzyme: | Bcl-2-related protein A1 | |
| inhibitor: | BDBM39149 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Bfl-1 assay materials: 1)Bfl-1 protein and FITC-Bid peptide (FITC-Ahx-EDIIRNIARHLAQVGDSMDR ) were obtained from Prof. John Reed (Sanford-Burnham Medical Research Institute, San Diego, CA) 2)Tb-anti-GST Antibody (Invitrogen, PV4216) 3)Assay Buffer: 25 mM Bis-Tris, pH 7.0, 1 mM TCEP, 0.005% Tween 20. 4)Bfl-1 working solution contained 7.4 nM GST-Bfl-1 in assay buffer 5)FITC-Bid working solution contained 5.6 nM FITC-Bid in assay buffer 6)FITC-Bid/ Tb-Ab working solution contained 5.6 nM FITC-Bid and 2.5 nM Tb-Ab in assay buffer. Solution was prepared fresh and kept on ice prior to use. Bfl-1 dose-response screening protocol: 1)Dose-response curves contained 10 concentrations of compounds obtained using 2-fold serial dilution. Compounds were serially diluted in 100% DMSO, and then diluted with water to 10% final DMSO concentration. 4 uL of compounds in 10% DMSO were transferred into columns 3-22 of Greiner 384-well white small-volume plates (784075). Each compound concentration was ass | |
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