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Multiplexed Profiling of the Cellular Stress Response at Multiple Pathway Points
EP24022
Poster Title: Multiplexed Profiling of the Cellular Stress Response at Multiple Pathway Points
Submitted on 01 Mar 2016
Author(s): Brad Hook, Chris Eggers, Matthew Robers, Amy Landreman
Affiliations: Promega Corporation
This poster was presented at N/A
Poster Views: 1,275
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Poster Information
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.
Summary: This poster demonstrates the use of a new dual-luciferase reporter assay for profiling cellular stress in multiple pathway points in the same population of cells. The assay format is homogeneous or 'add-mix-add-mix-read'. The authors used hypoxic and antioxidant cell models..References: Report abuse »
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