Department of Chemistry, University of Washington, Seattle WA, USA.
Electrophoresis. 2010 Aug;31(15):2650-4. doi: 10.1002/elps.201000151.
CIEF and CZE are coupled with LIF detection to create an ultrasensitive 2-D separation method for proteins. In this method, two capillaries are joined through a buffer-filled interface. Separate power supplies control the potential at the injection end of the first capillary and at the interface; the detector is held at ground potential. Proteins are labeled with the fluorogenic reagent Chromeo P503, which preserves the isoelectric point of the labeled protein. The labeled proteins were mixed with ampholytes and injected into the first-dimension capillary. A focusing step was performed with the injection end of the capillary at high pH and the interface at low pH. To mobilize components, the interface was filled with a high pH buffer, which was compatible with the second-dimension separation. A fraction was transferred to the second-dimension capillary for separation. The process of fraction transfer and second dimension separation was repeated two dozen times. The separation produced a spot capacity of 125.
CIEF 和 CZE 与 LIF 检测相结合，创建了一种用于蛋白质的超灵敏二维分离方法。在该方法中，通过充满缓冲液的界面将两个毛细管连接在一起。单独的电源控制第一根毛细管的注入端和界面处的电势；检测器保持在接地电位。蛋白质用荧光试剂 Chromeo P503 进行标记，该试剂保留了标记蛋白质的等电点。标记的蛋白质与两性电解质混合并注入到第一维毛细管中。通过将毛细管的注入端置于高 pH 值处并将界面置于低 pH 值处来进行聚焦步骤。为了使组分迁移，界面充满了高 pH 值缓冲液，该缓冲液与二维分离兼容。将一部分转移到二维毛细管中进行分离。分馏转移和二维分离的过程重复了二十几次。该分离产生了 125 个斑点容量。
