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Simple Western analysis of NFκB signaling cascade proteins
Author(s) -
Boge Annegret,
Nguyen Uyen,
Ramirez Francisco,
Xu Hui,
Kazakova Irina,
Yang Tom,
Gavin Robert
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.lb172
Subject(s) - signal transduction , blot , biomarker , computational biology , nf κb , simple (philosophy) , biology , cancer research , chemistry , microbiology and biotechnology , genetics , gene , philosophy , epistemology
Aberrant expression in the NFκB pathway is a common occurrence in inflammatory and stress‐induced diseases such as cancer. Understanding how NFκB signaling impacts disease progression is important to the development of novel therapeutics. Western blots, while semi‐quantitative and tedious to perform are commonly used. The Simple Western™ platform, provides a solution that enables the complete automation of traditional Westerns allowing analysis of up to 96 samples in a single experiment. Simple Westerns use nanoliter volumes, enabling the characterization of precious or limited quantity samples providing reproducible and quantitative results. Targets from the NFκB pathway, including IκB, NFκB subunits c‐ Rel, p65, p50/p105 along with loading controls were run on the Simple Western platform. In response to TNF‐α treatment, statistically significant changes of key target proteins were characterized. The Simple Western permits quantitative high throughput analysis of signaling pathways at a level of efficiency and reproducibility not obtainable by traditional Westerns, allowing better characterization of regulatory events and key biomarker targets involved in the progression of disease states.