Hermansen L F, Juul J, Sletten K
Department of Biochemistry, University of Oslo, Norway.
Electrophoresis. 1996 Apr;17(4):720-2. doi: 10.1002/elps.1150170417.
The applicability and reproducibility of a previously described (L. F. Hermansen et al., Electrophoresis 1993, 14, 1302-1306) centrifuge-blotting procedure for capturing subnanomolar amounts of protein on polyvinylidene difluoride membranes for direct Edman degradation was further investigated. Proteins with different molecular masses were centrifuge-blotted onto Immobilon CD membranes. Simultaneous blotting and desalting was achieved with an overall yield of 15-56% after 2 h centrifugation for proteins with a molecular mass of 12-30 kDa. Centrifugation of myoglobin for 6 h resulted in an overall yield of 72%. The subnanomolar amounts obtained were also sufficient to conduct cyanogen bromide cleavage in situ on proteins with blocked NH2-terminus and to generate sequence information.
先前已描述过的（L. F. 赫尔曼森等人，《电泳》，1993年，第14卷，第1302 - 1306页）用于在聚偏二氟乙烯膜上捕获亚纳摩尔量蛋白质以进行直接埃德曼降解的离心印迹法的适用性和可重复性得到了进一步研究。不同分子量的蛋白质被离心印迹到Immobilon CD膜上。对于分子量为12 - 30 kDa的蛋白质，在2小时离心后实现了同时印迹和脱盐，总产率为15 - 56%。肌红蛋白离心6小时的总产率为72%。所获得的亚纳摩尔量也足以对氨基末端被封闭的蛋白质进行原位溴化氰裂解并生成序列信息。
