| Solution Information | help | |
| Enzyme: | Glutathione S-transferase Mu 1 | |
| inhibitor: | BDBM47553 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | In this counterscreen assay, compounds were evaluated for the ability to interfere with the binding of the GST fusion proteins to the GSH beads (thus producing potential false positive results). Dose response experiments were constructed in which compounds at 10 mM concentration in DMSO were serially diluted 1:3.16 eight times for a total of nine different test compound concentrations. Final compound dilutions in DMSO ranged from 1 microM to 10 mM. These dilutions were then further diluted 1 to 100 to give an assay concentration range of 10 nanoM to 100 microM. GSH beads were incubated overnight with 50 nM GST-green fluorescent protein similar to conditions in the in the screening assay. Sample acquisition and preliminary analysis was conducted with the HyperCyt(R) high throughput flow cytometry platform. The HyperCyt system interfaces a flow cytometer and autosampler for high-throughput microliter-volume sampling from 384-well microtiter plates. The stream of particles is exci | |
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