Lam Hoang-Trang, Antonioli Paolo, Righetti Pier Giorgio, Citterio Attilio, Girault Hubert
Laboratoire d'Electrochimie Physique et Analytique, EPFL SB ISIC LEPA, Lausanne, Switzerland.
Electrophoresis. 2007 Jun;28(12):1860-6. doi: 10.1002/elps.200600763.
A minidevice for performing gel-free proteome prefractionation via conventional IEF in soluble carrier ampholyte buffers is reported here. It consists of a compact block of polyoxymethylene in which eight samples and two electrode chambers are machined. Each of the eight sample chambers can be filled with up to 120 microL of sample and has the following size: 7 mm width, 3 mm depth and 10 mm height. The anodic and cathodic compartments have the same width and height as the sample chambers, but with a depth of 6 mm, thus accepting up to 250 microL of electrodic solutions. Focusing is in general accomplished in 2 h with a voltage gradient of up to 1000 V (7 cm electrode distance). Easy fractionation and collection of the content of the eight chambers is achieved by simply pressing a rubber diaphragm against the edges of the thin walls separating each well, this automatically breaking liquid continuity. The performance of this device has been tested by subfractionating total cell lysates of a human cancer cell line (U2Os) and of Escherichia coli bacterial cells, and by analysing the content of each chamber by mono-dimensional SDS-PAGE and 2-D maps.
本文报道了一种通过在可溶性载体两性电解质缓冲液中进行常规等电聚焦（IEF）来实现无凝胶蛋白质组预分级分离的微型装置。它由一块紧凑的聚甲醛块组成，其中加工有八个样品室和两个电极室。八个样品室中的每一个最多可容纳120微升样品，其尺寸如下：宽7毫米、深3毫米、高10毫米。阳极和阴极隔室的宽度和高度与样品室相同，但深度为6毫米，因此最多可容纳250微升电极溶液。一般在2小时内完成聚焦，电压梯度高达1000伏（电极距离7厘米）。通过简单地将橡胶隔膜压在分隔每个孔的薄壁边缘上，就可以轻松地对八个室的内容物进行分级分离和收集，这会自动打破液体连续性。通过对人癌细胞系（U2Os）和大肠杆菌细菌细胞的总细胞裂解物进行亚分级分离，并通过一维SDS-PAGE和二维图谱分析每个室的内容物，对该装置的性能进行了测试。
