Nishino H, Ishibashi T, Nakamura H
Department of Biochemistry, Hokkaido University School of Medicine, Sapporo, Japan.
Biochem Mol Biol Int. 1994 Sep;34(2):409-17.
A simple method for preparation of highly-purified antibodies from antiserum was developed by using antigen-blotted polyvinylidene difluoride transfer membrane as an affinity matrix: The antigen was purified by SDS-polyacrylamide slabgel electrophoresis. Purified antibody preparations, which had been eluted from the Gly-HCl buffer (pH 2.5)-pretreated affinity matrix, by treatment with 1,4-dioxane, were more effective to obtain specifically stained band in immunoblot analysis than antiserum or immunoglobulin fractions purified by already reported methods. Antigen-blotted membranes were repeatedly available without remarkable release of antigen or decrease of antigenicity. This method was also applied to the examination of cross-reacting proteins with rat liver fatty acid binding protein in the cytosol fraction.
通过使用抗原印迹的聚偏二氟乙烯转移膜作为亲和基质，开发了一种从抗血清中制备高纯度抗体的简单方法：通过SDS-聚丙烯酰胺平板凝胶电泳纯化抗原。从经甘氨酸-盐酸缓冲液（pH 2.5）预处理的亲和基质上洗脱的纯化抗体制剂，经1,4-二氧六环处理后，在免疫印迹分析中比通过已报道方法纯化的抗血清或免疫球蛋白组分更有效地获得特异性染色带。抗原印迹膜可重复使用，而不会有明显的抗原释放或抗原性降低。该方法还应用于检测细胞质部分中与大鼠肝脏脂肪酸结合蛋白发生交叉反应的蛋白质。
