求问 2Xlaemmli sample buffer 怎么配制？

阿颠

2013-08-31

我最近要做 pulldown 分析，当珠子沉下后要用 2Xlaemmli sample buffer 洗脱，但是我不知道 2Xlaemmli sample buffer 怎么配制，求高手指导。2Xlaemmli sample buffer 与2Xloading buffer有什么区别！！还有 DDT 可以替代巯基丁醇吗？如果可以其中的量应该怎么替换呢？

1 个回答

liona_lin

2013-09-07

有帮助

1. Sample Buffer, Laemmli 2× Concentrate:Solution contains 4% SDS, 20% glycerol, 10% 2-mercaptoethanol, 0.004% bromphenol blue and 0.125 M Tris HCl, pH approx. 6.8. 2. SDS gel-loading buffer (2X) 100 mM Tris-Cl (pH 6.8), 4% (w/v) SDS (sodium dodecyl sulfate; electrophoresis grade), 0.2% (w/v) bromophenol blue, 20% (v/v) glycerol, 200 mM DTT (dithiothreitol)Store the SDS gel-loading buffer without DTT at room temperature. Add DTT from a 1 M stock just before th