| Assay Method Information | |
| | Luminescence Cell-Based Dose Confimation HTS to Identify Inhibitors of Heat Shock Factor 1 (HSF1) |
| Description: | Keywords: Heat Shock Factor-1 (HSF-1), Stress Response, MG132, NIH3T3, Luminescence Assay Overview: Modified NIH3T3, transformed to express firefly luciferase under the control of a HSF-1 response element, will be exposed to small molecules. After 30min exposure to small molecules, proteasome inhibitor MG132 is added to elicit a stress response. After 8hr incubation in the presence of this stressor, the amount of HSF-1 mediated luciferase expression is measured using a luminescence detection reagent. Expected Outcome: Identification of HSF-1 inhibitors in those instances where there is a loss of luminescence signal due to the prevention of HSF-1 being able to drive the expression of the luciferase reporter. Potential false positives in this assay included those compounds which act not by specifically inhibiting HSF-1, but by inhibiting luciferase or are broad spectrum inhibitors of transcription/translation. |
| Affinity data for this assay | |
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