Ink-native electrophoresis: An alternative to blue-native electrophoresis more suitable for in-gel detection of enzymatic activity.
Ink-native electrophoresis: An alternative to blue-native electrophoresis more suitable for in-gel detection of enzymatic activity.
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墨水原生电泳：蓝色原生电泳的替代方案，更适合酶活性的凝胶内检测。
DOI：
10.1016/j.ab.2013.05.026
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Ishida A,
中科院分区：
文献类型：
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作者：
Kaneko K;Sueyoshi N;Kameshita I;Ishida A,
蓝色天然电泳（BNE）是分析蛋白质复合物的一种有用技术，但BNE中使用的考马斯亮蓝（CBB）染料经常妨碍酶活性的凝胶内检测。在这里，我们报告了一种改进的方法，称为墨水自然电泳（INE），其中Pelikan 4001钢笔墨水作为电荷转移剂，而不是CBB。INE比BNE更适合于聚丙烯酰胺凝胶电泳（PAGE）后的蛋白激酶活性的胶内检测，并且其在蛋白复合物分离方面的性能与常规BNE相当。INE为分离和分析各种蛋白质复合物提供了有力的工具。
Blue-native electrophoresis (BNE) is a useful technique for analyzing protein complexes, but the Coomassie brilliant blue (CBB) dye used in BNE often hampers in-gel detection of enzymatic activity. Here we report an improved method, termed ink-native electrophoresis (INE), in which Pelikan 4001 fountain pen ink is used as a charge-shifting agent instead of CBB. INE is more suitable than BNE for in-gel detection of protein kinase activity after polyacrylamide gel electrophoresis (PAGE), and its performance in protein complex separation is comparable to that of conventional BNE. INE may provide a powerful tool to isolate and analyze various protein complexes.