Fahnestock S R, Strycharz W A, Marquis D M
J Biol Chem. 1981 Oct 10;256(19):10111-6.
Antibodies prepared against individual 50 S ribosomal subunit proteins from Escherichia coli were reacted with 70 S ribosomal proteins from Bacillus stearothermophilus in order to identify homologous protein pairs. B. stearothermophilus proteins were separated by two-dimensional polyacrylamide gel electrophoresis and transferred electrophoretically to diazobenzyloxymethyl paper to which they became covalently attached. The paper was then washed with antiserum followed by radioactive protein A, and the resulting antigen-antibody-protein A complexes were located by autoradiography. Seventeen cross-reacting protein pairs were identified.
制备了针对大肠杆菌单个50 S核糖体亚基蛋白的抗体,并使其与嗜热脂肪芽孢杆菌的70 S核糖体蛋白反应,以鉴定同源蛋白对。嗜热脂肪芽孢杆菌的蛋白通过二维聚丙烯酰胺凝胶电泳进行分离,然后通过电泳转移至重氮苄氧基甲基纸上,并与之共价结合。接着用抗血清洗涤该纸,随后加入放射性蛋白A,通过放射自显影定位产生的抗原-抗体-蛋白A复合物。鉴定出了17对交叉反应蛋白对。