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Multiplexed Profiling of the Cellular Stress Response at Multiple Pathway Points
EP24022
Multiplexed Profiling of the Cellular Stress Response at Multiple Pathway Points
Submitted on 01 Mar 2016

Brad Hook, Chris Eggers, Matthew Robers, Amy Landreman
Promega Corporation
This poster was presented at N/A
Poster Views: 1,080
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Poster Abstract
Many cellular responses to changing environmental conditions occur at multiple points in the signaling pathway in order to allow for cellular adaptation and survival. The cellular stress response often involves changes in the stability of a critical regulatory protein followed by downstream modulation of specific transcriptional targets. These changes can occur at sub-cytotoxic exposure levels providing predictive measures of toxicity. Irregular modulation of these pathways can also occur in tumor environments or disease states resulting in undesirable outcomes, presenting potential drug targets.

Dual-reporter assays provide a way to monitor multiple pathway points in the same population of cells. The Nano-Glo Dual-luciferase (NanoDLR) assay allows for sequential monitoring of firefly luciferase (Fluc) and Nanoluc luciferase (Nluc) from the same sample using a simple add-read-add-read protocol. Efficient quenching of the initial Fluc read coupled with the bright Nluc signal give high sensitivity to both reporters. In addition, the NanoDLR assay can be coupled with a fluorescent cell viability assay allowing for a more complete profile of the overall cellular response to be assayed from a single sample.


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