抗猪脂肪细胞膜蛋白抗体的制备及测定

Preparation and identification of antibodies against porcine adipocyte plasma membrane protein

猪屠宰后,立刻取其背部皮下脂肪组织在37℃膜萃取液中匀浆,用差速离心法和密度梯度离心法提取脂肪细胞膜蛋白,经SDS-PAGE分析,背部脂肪细胞膜蛋白与其他组织细胞膜蛋白具有较大差异,但也有一些相同的膜蛋白.以猪背部脂肪细胞膜蛋白为抗原,免疫雄性家兔,制备抗猪脂肪细胞膜蛋白血清,经酶联免疫反应(ELISA)测定,血清效价达到1∶12800.同样,用ELISA和Western-blotting测定抗体与其他组织细胞膜的交叉反应,结果显示:抗猪脂肪细胞膜抗体与其他组织细胞膜有交叉反应,但反应性不高.

The adipose tissue extracted from crossbred pigs was homogenized in 37℃ membrane extractant. Membrane proteins of pig subcutaneous adipocyte and other tissue cells were extracted by differential centrifugation method and density gradient centrifugation method. Comparative analyses of membrane protein profiles between adipoeyte and other tissue cell proteins was conducted by SDS-PAGE. Though componets of membrane proteins were different among pig subcutaneous and other tissue cells, some membrane proteins were identical. The antibodies against porcine adipoeyte plasma membrane protein were produced by immunising with subcutaneous adipocyte plasma membrane protein as antigen, and its titer reached 1：12800 detected by ELISA. Likewise low cross-reactivity of the antisera to other pig tissues could be detected by ELISA and Western-bohting.