摘要
用人工合成的叶绿体细胞色素b_6-f复合物亚单位Ⅳ(17kD)中3个含10个氨基酸残基的小肽,分别与卵清蛋白偶联,经高压液相层析纯化后作为抗原去免疫家兔。获得抗17kD的3种单克隆抗体。蛋白质印渍术结果表明3个抗体均与17kD有免疫亲和反应。酶联免疫吸附测定表明,3种抗体均与菠菜类囊体膜有免疫亲和反应。因此这些抗体可用来作探测17kD在类囊体膜内、外两侧的分布及定位的探针。
Three synthetic decapeptides of subunit Ⅳ of cytochrome b(?)-f complex were coulpled respectively with ovalumine and purified using High-Performance Liquid Chromatography. They were served as antigens and injected into rabbits. Five weeks later, three kinds of monoclonal antibodies were obtained. Western blotting showed that there were immunoaffinity reactions between and 17 kD-1, anti 17 kD-2, anti 17 kD-3 and 17kD band of cytochrome b(?)-f complex from spinach chloroplasts. Enzyme-linked immunosorbent assay showed that there were immunoaffinity reactions between anti 17 kD-1, anti 17 kD-2, anti 17 kD-3 and thylakoid membranes from spinach chloroplasts. Therefore, these antibodies can be used as probes to survey the location and orientation of 17 kD on the two sides of thylakoid membranes.
关键词
小肽
单克隆抗体
叶绿体
细胞色素
Shortpeptide
Monoclonal antibody
Probe
Subunit Ⅳ of cytochrome b_6-f complex
Western blotting
Enzyme-linked immunosorbent assay
