Fries E
Biochim Biophys Acta. 1976 Dec 14;455(3):928-36. doi: 10.1016/0005-2736(76)90061-4.
The molecular weight of proteins in protein-detergent complexes can be determined from ultracentrifugation experiments if the amount of bound detergent is known. A new sensitive method to measure the binding of the nonionic detergent Triton X-100 to proteins has been developed. For the membrane proteins studied, less than 50 mug of protein was required to achieve an accuracy of 10% in the determination of the detergent-protein weight ratio. The proteins were equilibrated with the detergent by electrophoresis into polyacrylamide gels containing radioactively labelled Triton X-100. The gels were then sliced and the amount of bound detergent calculated from the increase in radio-activity in the slices containing the protein zone. The amounts of protein were determined by amino acid analysis of identical protein zones cut from gels running parallel.
如果已知结合去污剂的量，那么蛋白质 - 去污剂复合物中蛋白质的分子量可以通过超速离心实验来确定。一种用于测量非离子去污剂Triton X - 100与蛋白质结合的新的灵敏方法已经开发出来。对于所研究的膜蛋白，在确定去污剂 - 蛋白质重量比时，达到10%的准确度所需的蛋白量少于50微克。通过电泳将蛋白质与去污剂在含有放射性标记的Triton X - 100的聚丙烯酰胺凝胶中达到平衡。然后将凝胶切片，根据含有蛋白质区域的切片中放射性的增加来计算结合去污剂的量。蛋白质的量通过对与平行运行的凝胶上切下的相同蛋白质区域进行氨基酸分析来确定。
