Rabilloud T, Girardot V, Lawrence J J
Laboratoire de Biologie Moleculaire du Cycle Cellulaire, Grenoble.
Electrophoresis. 1996 Jan;17(1):67-73. doi: 10.1002/elps.1150170112.
We tested the recently introduced methylene blue-toluene sulfinate-diphenyl-iodonium polymerization system in order to prepare acetic acid-urea-Triton X-100 gels for histone separations. When compared to standard persulfate-based initiators, this system exhibited several advantages. First, the polymerization proceeds at a much faster rate but is easily controlled since it is light-dependent. Second, no prerunning of the gel was required, since there is no oxidizing molecule able to introduce artifacts. Moreover, this procedure produces gels presenting cleaner backgrounds with silver staining. The ability of the procedure to carry out high resolution two-dimensional electrophoresis of histones was also examined. High resolution two-dimensional gels, using photopolymerized acidic gels as the first or second dimension were obtained.
我们测试了最近引入的亚甲蓝-甲苯亚磺酸盐-二苯基碘鎓聚合体系，以制备用于组蛋白分离的乙酸-尿素-曲拉通X-100凝胶。与基于过硫酸盐的标准引发剂相比，该体系具有几个优点。首先，聚合反应进行得更快，但由于其依赖光，所以易于控制。其次，不需要对凝胶进行预电泳，因为不存在能够引入假象的氧化分子。此外，该方法制备的凝胶经银染后背景更清晰。我们还研究了该方法进行组蛋白高分辨率二维电泳的能力。使用光聚合酸性凝胶作为第一维或第二维，获得了高分辨率二维凝胶。
