| Solution Information | help | |
| Enzyme: | Nuclear receptor subfamily 4 group A member 1 | |
| inhibitor: | BDBM40726 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Bcl-B assay materials: 1) Bcl-B protein and FITC-TR3-R8 peptide (FITC-Ahx-FSRSLHSLL-GX-R8 ) were obtained from Prof. John Reed (Sanford-Burnham Medical Research Institute, San Diego, CA) 2) Assay buffer: 37.5 mM HEPES-NaOH, pH 7.5, 1.5 mM TCEP, 0.0075% Tween 20. 3) Bcl-B working solution contained 25 nM Bcl-B in assay buffer. Solution was prepared fresh and kept on ice prior to use. 4) Assay buffer with beta-GP: 37.5 mM HEPES-NaOH, pH 7.5, 30 mM beta-glycerophosphate (beta-GP), 1.5 mM TCEP, 0.0075% Tween 20. 5) FITC-TR3 working solution contained 50 nM FITC-TR3-R8 peptide in the assay buffer with beta-GP. Bcl-B HTS protocol: 1) 4 uL of 100 uM compounds in 10% DMSO were dispensed in columns 3-24 of Greiner 384-well black small-volume plates (784076). Columns 1 and 2 contained 4 uL of 10% DMSO. 2) Positive control wells, that contained no Bcl-B, were assigned to column 1. Assay buffer (8 uL) was added to these wells using the WellMate bulk dispenser (Matrix). 3) 8 uL of Bcl-B worki | |
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