Method for resolution and western blotting of very large proteins using agarose electrophoresis.
Method for resolution and western blotting of very large proteins using agarose electrophoresis.
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使用琼脂糖电泳对非常大的蛋白质进行分辨率和蛋白质印迹的方法。
DOI：
10.1007/978-1-4939-2694-7_30
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Warren,ChadM
中科院分区：
文献类型：
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作者：
Greaser,MarionL;Warren,ChadM
大于 200 kDa 的蛋白质很难使用聚丙烯酰胺凝胶进行电泳分离，并且它们在蛋白质印迹过程中的转移通常是不完整的。开发了垂直 SDS 琼脂糖凝胶系统，该系统大大提高了对非常大的蛋白质的解析能力。可以实现将肌联蛋白 (Mr 3,000–3,700 kDa) 这样大的蛋白质完全转移到印迹上。在上层储存缓冲液和转移缓冲液中添加巯基还原剂可显着改善大蛋白的印迹。
Proteins larger than 200 kDa are difficult to separate electrophoretically using polyacrylamide gels, and their transfer during western blotting is typically incomplete. A vertical SDS agarose gel system was developed that has vastly improved resolving power for very large proteins. Complete transfer of proteins as large as titin (Mr 3,000–3,700 kDa) onto blots can be achieved. The addition of a sulfhydryl reducing agent in the upper reservoir buffer and transfer buffer markedly improves the blotting of large proteins.