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(The FASEB Journal. 2008;22:645.16)
© 2008 FASEB
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(The FASEB Journal. 2008;22:645.16.)
© 2008 FASEB

645.16

Analysis of Signaling Pathways using Antibody Microarrays and RNA Interference

Janaki Narahari, Aric Morgan, Krishna Vattem, Yvonne Sullivan, Douglas Hughes, Eric Hommema, Greg Kilmer and Brian Webb

Research & Development, Thermo Fisher Scientific, Rockford, IL

ABSTRACT

The study of complex cellular signaling pathways requires powerful and specific tools for the manipulation and quantification of protein levels and activity. We have developed a new set of tools to streamline the experimental approach for pathway analysis. These include a series of validated Thermo Scientific SuperSignal® siRNA/Antibody Kits for monitoring gene-silencing at the protein level, slide-based antibody microarrays, and several kits for cell- based assays. We have demonstrated target-specific applications to confirm the functionality of the various kits. For example, the expression profile of various cytokines and MMPs were analyzed by protein microarrays following inhibition of several signaling pathways using RNA interference. Knockdown of NFB protein in A549 cells using siRNA resulted in significant inhibition of cytokine production. The activation of signaling proteins in these pathways was studied using our Thermo Scientific Cellomics® HCS Reagent Kits. Our results demonstrate the high quality and utility of these reagents for the efficient profiling of different pathways.





This Article
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Right arrow Articles by Webb, B.
PubMed
Right arrow Articles by Narahari, J.
Right arrow Articles by Webb, B.