Wenge Birger, Bönisch Heinz, Grabitzki Julia, Lochnit Günter, Schmitz Brigitte, Ahrend Michael H J
Institute of Pharmacology and Toxicology, University of Bonn, Bonn, Germany.
Electrophoresis. 2008 Apr;29(7):1511-7. doi: 10.1002/elps.200700546.
Due to their poor solubility during IEF membrane proteins cannot be separated and analyzed satisfactorily with classical 2-DE. A more efficient method for such hydrophobic proteins is the benzyldimethyl-n-hexadecylammonium chloride (16-BAC)/SDS-PAGE, but the corresponding protocol is intricate and time-consuming. We now developed an easy-to-handle electrophoresis method in connection with a novel device which enables reproducible separation of ionic solubilized membrane proteins using individually rehydrated plastic sheet gel strips. These strips are suitable for the first dimension in a 2-D 16-BAC/SDS system and can be handled easily; this is demonstrated by the separation of membrane proteins of human embryonic kidney (HEK293) cells.
由于膜蛋白在等电聚焦过程中溶解性较差，无法通过经典的双向电泳（2-DE）得到令人满意的分离和分析。对于这类疏水蛋白，一种更有效的方法是使用苄基二甲基正十六烷基氯化铵（16-BAC）/十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE），但相应的实验方案复杂且耗时。我们现在开发了一种易于操作的电泳方法，并结合一种新型装置，该装置能够使用单独复水的塑料片凝胶条对离子溶解的膜蛋白进行可重复的分离。这些条带适用于二维16-BAC/SDS系统的第一维，并且易于操作；人胚肾（HEK293）细胞膜蛋白的分离证明了这一点。
