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A Comparison of Western Blotting Techniques
Using Phospho-Specific Antibody Detection of Protein Kinase Activation
By Erik M. Schaefer and Bruce W. Jarvis
Promega Corporation
Western blotting is a popular method for using highly selective antibodies to
visualize proteins in cell or tissue extracts. Indeed, the availability of specific
antibodies that selectively target post-translational modifications (e.g.,
phosphorylation) of the protein of interest can provide a highly sensitive and
non-radioactive measure of protein kinase activation via Western analysis. In this article
the detection of MAP kinase activation using Promega's Anti-ACTIVETM
MAPK pAb and prequalified HRP- and AP-labeled secondary antibodies is presented to
illustrate commonly used Western methods.
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