Berntzen H B, Fagerhol M K
Institute of Clinical Biochemistry, Rikshospitalet, Oslo, Norway.
Scand J Clin Lab Invest. 1990 Nov;50(7):769-74. doi: 10.1080/00365519009091071.
L1 is a major granulocyte and monocyte protein with a Mr of 36.5 kDa. It is found mainly in the cytosol of these cells. Purified L1 is shown, on two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), to contain three subunits. In this study, 6 mol/l concentration of urea was found to be sufficient for disassembly of the polypeptides, and urea-containing preparative isoelectric focusing gel was used for separation of high quantities of the subunits. The pI of the eluted subunits were 5.8, 6.1 and 7.1. When tested on 2D-PAGE, the isolated subunits were found at their typical locations. Polyclonal rabbit antibodies were produced against the subunits, and the antisera were, on dot-blot, found to react with the different subunits as well as the purified L1.
L1是一种主要的粒细胞和单核细胞蛋白,分子量为36.5 kDa。它主要存在于这些细胞的胞质溶胶中。经二维聚丙烯酰胺凝胶电泳(2D-PAGE)显示,纯化的L1含有三个亚基。在本研究中,发现6 mol/l浓度的尿素足以使多肽解离,含尿素的制备性等电聚焦凝胶用于分离大量亚基。洗脱的亚基的等电点分别为5.8、6.1和7.1。在2D-PAGE上进行检测时,发现分离出的亚基位于其典型位置。制备了针对这些亚基的兔多克隆抗体,经斑点印迹法检测,发现抗血清能与不同亚基以及纯化的L1发生反应。