Unable to connect to database - 08:26:49 Unable to connect to database - 08:26:49 SQL Statement is null or not a SELECT - 08:26:49 SQL Statement is null or not a DELETE - 08:26:49 Botany & Plant Biology 2007 - Abstract Search
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Abstract Detail


Intracellular Signaling

Anderson, Jeffrey C. [1], Peck, Scott C. [2].

A simple and rapid technique for stoichiometric detection of protein phosphorylation.

Recent advances in phosphoproteomic-based approaches have rapidly increased the inventory of known phosphoproteins. However, most techniques for measuring the phosphorylation status of individual proteins remain technically demanding to perform (e.g. two-dimensional (2-D) gel electrophoresis), or require a priori knowledge of specific phosphorylation sites (e.g. immunoblot detection with phospho-specific antibodies). Furthermore, although phosphorylation can alter the migration of proteins in SDS-PAGE gels, this shift only occurs for a subset of phosphoproteins. To overcome these limitations, we developed a method for detecting phosphorylation that involves isoelectric focusing of proteins in a vertical mini-gel format under denaturing conditions, followed by immunoblot detection of the protein of interest. The method is simple, easily implemented with standard laboratory equipment, and provides a rapid means to analyze the phosphorylation status of individual proteins. In addition, this approach provides a stoichiometric measure of phosphorylation making it possible to determine the extent to which a protein is phosphorylated. Examples of the implementation of this technique will be presented.


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1 - University of Missouri, Biochemistry, Christopher Bond Life Sciences Center, 1201 Rollins St., Columbia, MO, 65211, USA
2 - University of Missouri, Biochemistry

Keywords:
Phosphorylation
protein post-translational modification
kinase.

Presentation Type: Plant Biology Abstract
Session: P
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM
Number: P33024
Abstract ID:2294


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