HSF1-related Assays (Heat shock)
The LightSwitch™ Validated Heat Shock (HSF) Pathway Collection is a set of sequence-verified, transfection-ready, promoter reporter constructs that can be used to accurately quantify transcriptional activation in response to induction of the heat shock pathway. These constructs are a subset of the 18,000-member LightSwitch Promoter Reporter GoClone™ Collection that were selected based on sequence motif analysis, published information and in-house analysis of each promoter's activity* using our LightSwitch Luciferase Assay. The Heat Shock Biomarker Set contains 8 of the highest responding promoters under our chosen test conditions, as well as the promoters of 2 housekeeping genes (Figure 1). Active Motif also offers a HSF Pathway Reporter Stable Cell Line to facilitate high-throughput screening of the heat shock pathway.
Figure 1: Induction ratios for each member of the Heat Shock Biomarker Set.
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Heat Shock (HSF) Pathway
The regulation of heat-shock genes is critical in cell stress response and the maintenance of cellular homeostasis. Heat-shock gene transcription is regulated by HSF (heat shock factor) transcription factors in response to various types of environmental stress such as heat, starvation, hypoxia, infection and toxin exposure. Heat shock proteins (HSPs) are also well-characterized for their roles as chaperones for protein synthesis, assembly, secretion and degradation. As specific HSPs have been implicated as central players in apoptosis, tumor survival, and immune response, understanding the regulation of the heat shock response is critical.
The collection of validated LightSwitch heat shock-related promoter reporters enables you to:
- Understand the mechanisms by which heat shock regulated genes are induced or repressed
- Quantify the functional consequences of transcription factor binding – while many other technologies give a qualitative view of transcription factor level, our promoter reporter assays quantify the effects of transcription factor binding
- Confirm data from ChIP, ChIP-chip or ChIP-Seq experiments
- Measure the effect of sequence variants and mutagenesis on promoter function
- Screen for promoter activation or develop activity profiles across the heat shock response pathway for many compounds or conditions in parallel
LightSwitch Luciferase Assay System
The LightSwitch Luciferase Assay System is a complete solution for studying transcriptional & post-transcriptional gene regulation in living mammalian cells. It enables you to directly measure the functional activity of promoters and 3´UTRs through use of an engineered luciferase gene (RenSP) and optimized assay & transfection reagents that ensure superior, reproducible results. In addition to Validated Pathway Collections, the LightSwitch System includes collections of over 30,000 cloned human promoter reporter constructs and human 3´UTR reporter constructs, miRNA mimics and inhibitors, stable cell lines, as well as collections of synthetic long-range response element constructs and synthetic 3´UTRs target validation constructs that can provide more sensitivity than endogenous sequences. Comprehensive Custom Services are also available
LightSwitch Promoter Reporter Assays.
Advantages of the LightSwitch Luciferase Assay System
- Quantitative – Novel RenSP luciferase technology allows you to measure promoter activity with industry-leading sensitivity and dynamic range.
- Simple, fast, complete solution – With pre-cloned LightSwitch Reporter vectors and optimized transfection & assay reagents, you can study regulation of your gene today. No cloning, DNA preparation or optimization is needed, and most studies do not require any internal transfection controls.
- Comprehensive and verified – The genome-wide LightSwitch Reporter Collections are sequence-verified, transfection-ready promoter and 3´UTR reporter vectors.
- Functionally insightful – Learn about the actual effects of transcription factor binding to verify computational predictions and supplement microarray or sequencing data.
- Cost-effective – Efficiently screen for activation and/or repression using a multitude of conditions.
The Heat Shock Biomarker Set contains 5 µg aliquots each of 8 promoter reporter constructs and 2 housekeeping promoter constructs (ACTB & RPL10) for use as controls. Click on the links below for clone information.