Heat Shock (HSF) Pathway Products

Introduction

SwitchGear Genomics offers transfection-ready reporter constructs to allow you to measure Heat Shock pathway activation. The SwitchGear Heat Shock promoter targets were picked from our genome-wide promoter collection based on sequence motif analysis, expression studies, and transcription factor binding data. We conducted experiments to generate a profile across the set of constructs with and without Heat Shock activation and offer a set of key responder constructs that show a strong induction response that can be used as biomarkers in screening applications.

Heat Shock Biomarker Set – 8 promoters, 2 controls

The selected constructs shown in the graph are 8 of the highest responding promoters under our chosen test conditions. Transient transfection assays were conducted in HT1080 cells in 96-well white plates. First, 5,000 cells per well were seeded in standard culture medium for 24 hours. Next, 50 ng of reporter plasmid DNA was transfected with Fugene-6 transfection reagent according to FuGENE standard protocols. After 12 hrs, three replicates of each transfected construct was moved to 43 degrees C, and three replicates were kept at 37 degrees C. Cells were incubated for 8 hours at their respective temperature. After 8 hrs, 100uL of luciferase assay reagent was added into each well, incubated at room temperature for 30 minutes, and then read in a standard plate luminometer. Each treatment for each reporter construct was assayed in triplicate, and the average of the 3 replicates without treatment and the average of the 3 replicates with treatment were recorded.