| Autor: |
Buehl, Christopher J., Deng, Xiexiong, Liu, Mengyu, McAndrew, Michael J, Hovde, Stacy, Xu, Xinjing, Kuo, Min-Hao |
| Zdroj: |
Biotechniques; August 2014, Vol. 57 Issue: 2 p72-80, 9p |
| Abstrakt: |
Protein acetylation and phosphorylation are key modifications that regulate both normal and pathological protein functions. The gel systems currently used for analyzing modified proteins require either expensive reagents or time-consuming second dimension electrophoresis. Here we present a neutral pH gel system that allows the analysis of acetylated and phosphorylated proteins. The neutral pH urea Triton-polyacrylamide gel electrophoresis (NUT-PAGE) system separates proteins based on their charge at pH 7.0 and generates discrete bands from each acetylated and/or phosphorylated species. In addition, the gel is composed of common and inexpensive laboratory reagents and requires only a single dimension of electrophoresis. We demonstrate the effectiveness of this system by analyzing the phosphorylated species of an acidic protein, α-synuclein, and both acetylated and phosphorylated species of a basic protein, histone H3. NUT-PAGE thus provides a cost-effective alternative for resolving acetylated and phosphorylated proteins, and potentially proteins with other post-translational modifications that alter net charge. |
| Databáze: |
Supplemental Index |
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