| Solution Information | help | |
| Enzyme: | Zinc finger protein mex-5 | |
| inhibitor: | BDBM49122 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | 2024-01-W01-03 7.5 nL of compound per well was pre-dispensed into black Aurora 1536 low-base plates (PN 00029844) in duplicate assay plates. Predispensed plates were stored lidded at room temperature for up to two weeks. For higher concentrations in dose retests, 15 or 30 nL was predispensed. MEX-5 protein, expressed in E. coli JM109 with a C-terminal fusion to maltose binding protein, was prepared by the assay provider at approximately 50 uM as previously described. A protein master mix was prepared at 200 nM in 1x buffer (20 mM tris pH 8.0 (Ambion), 40 mM NaCl (Ambion), 0.004% IPEGAL (Sigma), 100 uM zinc acetate dihydrate (Fluka)). The solution was loaded on the dispenser (Combi nL, Thermo) at room temperature. The target binding sequence TCR2 is a Temporal Control Region in the 3' untranslated region of the C. elegans gene glp-1 (abnormal Germ Line Proliferation). Fluorescent TCR2 RNA (5'-UUUCUUUAUAACUUGUUACAAUUUUUGAAA-FITC-3', Dharmacon / Thermo) mastermix was p | |
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