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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 small peptides with 10 amino acid residues in the synthetic chloroplast cytochrome b_6-f complex subunit Ⅳ (17kD) were respectively coupled with ovalbumin and purified by high pressure liquid chromatography as antigen Immunize the rabbit. Three anti-17kD monoclonal antibodies were obtained. Western blotting results showed that all three antibodies were immunoreactive with 17kD. Enzyme-linked immunosorbent assay showed that all three antibodies had immunological affinity with spinach thylakoid membranes. Therefore, these antibodies can be used as probes to detect the distribution and localization of 17kD in and around the thylakoid membrane.