| Solution Information | help | |
| Enzyme: | Signal transducer and activator of transcription 1-alpha/beta | |
| inhibitor: | BDBM43676 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay Overview: The purpose of this assay is to determine dose response curves for powder samples of compounds identified as possible STAT1 activator probes. In this assay STAT1 activation was measured using a murine NIH 3T3 fibroblast cell line that stably expresses a STAT1::luciferase construct. Test compounds were screened for their ability to increase IFN-gamma-mediated STAT1::luciferase reporter activity. Cells were exposed to test compound, followed by treatment with IFN-gamma. Changes in STAT1::luciferase activity were monitored by measuring luminescence. As designed, a STAT1 activator will increase IFN-gamma-mediated STAT1 transcription, thus increasing transcription of the luciferase reporter gene, leading to increased well luminescence. Compounds were tested in quadruplicates using a 10-point, 1:3 dilution series, starting at a nominal test concentration of 55.7 micromolar. Protocol Summary: NIH 3T3 cells were grown in T-175 flasks in Dulbecco's Modified Eagle's M | |
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