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用氨基丁基乙基异鲁米诺(ABEI)通过碳二亚胺(EDC)法标记分离的无载脂蛋白E的HDL_3。其标记率为0.89m mol/m mol HDL_3。HDL_3-ABEI与Apo A-Ⅰ和Apo A-Ⅱ抗体作用仍有免疫反应,说明标记后的HDL_3其免疫原性没有丧失。用HDL_3-ABEI与传代培养的正常人皮肤纤维母细胞作用,测其HDL受体活性。标记配基HDL_3-ABEI与人皮肤纤维母细胞HDL受体结合的最适量为20μg HDL_3-ABEI的蛋白质;人皮肤纤维母细胞HDL受体与HDL_3-ABEI在4℃结合的最适时间为1h;HDL_3-ABEI与人皮肤纤维母细胞结合时,人皮肤纤维母细胞最适量为0.1×10~6个细胞;未标记的HDL_3封闭结合HDL受体的最适用量为400μg。
Isolated HDL_3 of apolipoprotein E isolated by carbodiimide (EDC) method with aminobutylethyl isoluminol (ABEI). The labeling rate was 0.89m mol / m mol HDL_3. The effect of HDL_3-ABEI on Apo A-Ⅰ and Apo A-Ⅱantibodies was still immunoreactive, indicating that the labeled HDL_3 had no loss of immunogenicity. Using HDL_3-ABEI and sub-cultured normal human skin fibroblasts, measured HDL receptor activity. The optimal amount of HDL_3-ABEI labeled HDL_3-ABEI conjugated with human skin fibroblast HDL receptor was 20μg HDL_3-ABEI. The optimum time for HDL_3-ABEI binding to human HDL_3-ABEI was 1h. HDL_3-ABEI combined with human skin fibroblasts, the optimal amount of human skin fibroblasts 0.1 × 10 ~ 6 cells; HDL_3 unlabeled and HDL receptor block the optimal amount of 400μg.