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Low‐abundant protein using a Western blot enhancer
Author(s) -
Ganapathy Ramesh,
Nlend Marie,
Desai Surbhi,
Shiflett Steve,
Antharavally Babu,
Haney Paul
Publication year - 2011
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.25.1_supplement.lb105
Subject(s) - blot , western blot , enhancer , microbiology and biotechnology , chromogenic , biology , chemistry , biochemistry , gene expression , chromatography , gene
Western blotting is the standard method for protein identification; however, the procedure has many variables that can produce inconsistent results. Conventional Western blotting frequently results in low signal, poor sensitivity, sub‐optimal specificity and high background, which can mask target detection. The Thermo Scientific SuperSignal Western Blot Enhancer increases both signal intensity and sensitivity 3‐ to 10‐fold. We used the Western blot enhancer to detect GSK3 alpha and CDK5, which are typically undetectable using conventional chemiluminescent Western blotting. We were also able to decrease background when using the enhancer to detect ERK1 from K562 cell lysate. In addition to using the SuperSignal® Western Blotting Enhancer with chemiluminescent substrates, we present results of its effectiveness for chromogenic detection of beta‐catenin and fluorescent detection of MAP kinase in HeLa cell lysates.