Horvath A, Riezman H
University of Basel, Biochemistry Department, Switzerland.
Yeast. 1994 Oct;10(10):1305-10. doi: 10.1002/yea.320101007.
We have developed and evaluated an easy and rapid method for extraction of proteins from yeast cells for sodium dodecyl sulphate (SDS)-gel electrophoresis and Western blotting. The procedure comprises a centrifugation step to harvest the cells, addition of a sample buffer and heating, then another centrifugation step before applying the extracted proteins found in the supernatant to an SDS gel. It is applicable to the study of large numbers of samples in 1 day. This procedure is easier, quicker, and as efficient as procedures using base and 2-mercaptoethanol, but somewhat less efficient than lysis with glass beads under certain conditions.
我们开发并评估了一种简单快速的从酵母细胞中提取蛋白质的方法,用于十二烷基硫酸钠(SDS)-凝胶电泳和蛋白质免疫印迹分析。该方法包括一个用于收集细胞的离心步骤,加入样品缓冲液并加热,然后在将上清液中提取的蛋白质应用于SDS凝胶之前进行另一个离心步骤。它适用于在一天内研究大量样品。该方法比使用碱和2-巯基乙醇的方法更简便、快速且高效,但在某些条件下比用玻璃珠裂解的方法效率略低。