| Solution Information | help | |
| Enzyme: | E3 ubiquitin-protein ligase XIAP | |
| inhibitor: | BDBM44360 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | BIR3 assay materials: 1) Bir3 protein was provided by Prof. John C. Reed (Sanford-Burnham Medical Research Institute, San Diego, CA) and SMAC-rhodamine peptide (AVPIAQK-rhodamine) was provided by Richard Houghten (Torrey Pines Institute for Molecular Studies, San Diego, CA). 2) Assay buffer: 33.35 Hepes @ pH 7.5, 1.33 mM TCEP, 0.00667 % Tween 20, 0.265 uM BIR3 and 0.0267 uM SMAC-rhodamine. BIR3 protocol: 1) 15 ul of assay buffer was added to a Molecular Devices black 96 well plate (number 42-000-0117) in columns 1 through 10. 2) Five ul of compounds were added to the plate in columns 1 through 10 with a 16 point curve generated by 2 fold dilutions of each compound. Final compound concentration in the assay ranged from 100 uM to 0.00612 uM via two fold dilutions starting with 400 uM maximum concentration. 3) Positive controls without protein were in column 12. Negative controls with protein were in column 11. 4) The AVPF peptide was present with the same concentration range as t | |
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