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Updated: May 31, 2026

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach
Published on: February 21, 2019
A modified immunoblot method to identify substrates of protein kinases
Choong-Min Kang1, Wan Jin Jahng, Robert N Husson
1Department of Biological Science, Wayne State University, 5047 Gullen Mall, Detroit, MI 48202, USA. cmkang1@biology.biosci.wayne.edu
Abstract:
While protein kinases are key components in multiple cellular processes, efficient identification of cognate in vivo substrates remains challenging. Here we describe a powerful method to screen potential substrates of protein kinases by partial transfer of proteins from a 2D-PAGE gel to a Western blot membrane. This approach allowed precise pinpointing of candidate substrate spots in the 2D gel, and identifying physiological substrates of protein kinases in Mycobacterium tuberculosis.
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Western Blotting
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
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