Abstract
One dimensional polyacrylamide gel electrophoresis has been successfully used to detect protein phosphorylation. This method is very simple and highly reproducible. Hyperhosphorylated proteins usually migrate slowlier than dephosphorylated proteins. However, not all phosphorylated proteins can be readily detected, due to sub-optimal sample preparation and electrophoresis conditions. Here, an improved method is described that can detect phosphorylation of yeast proteins ranging from 15 kD to 200 kD. The improvement in gel electrophoresis should also be applicable to mammalian culture cells.
Keywords: Phosphorylation, Yeast, Mobility shift, Acrylamide gel, Western blot
Materials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
This protocol was adapted from and used in Wei and Zheng (2009) and Wei et al. (2009).
References
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