Assay Method Information

Assay Name:  Luminescence Cell-Based Dose Retest to Identify Potentiators of Heat Shock Factor 1 (HSF1)
Description:  Keywords: Heat Shock Factor-1 (HSF-1), Stress Response, MG132, NIH3T3, Luminescence Assay Overview: Confirmation testing of small molecules identified as increasing expression of a firefly luciferase under the control of a HSF-1 response element in modified NIH3T3 cells. For consistency with the primary assay in which these compounds were identified, after 30min exposure to small molecules, proteasome inhibitor MG132 is added to elicit a stress response. After an additional 8hr incubation, the amount of HSF-1 mediated luciferase expression is measured using a luminescence detection reagent. Expected Outcome: Identification of HSF-1 inducers in those instances where there is an increase of luminescence signal due to the enhancement 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 inducing HSF-1, but by stabilizing luciferase or are broad spectrum inducers of transcri
Affinity data for this assay

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