Gulle H, Brimacombe R, Stöffler-Meilicke M, Stöffler G
Max-Planck-Institut für Molekulare Genetik, Abteilung Wittmann, Berlin-Dahlem, Germany.
J Immunol Methods. 1987 Sep 24;102(2):183-6. doi: 10.1016/0022-1759(87)90075-5.
A method is described for the rapid immunological identification of proteins in studies of multicomponent systems. In this case the system is the E. coli ribosome, and the ribosomal proteins to be identified are covalently attached to fragments of labelled ribosomal RNA as a result of chemical cross-linking procedures. Antisera raised against the individual ribosomal proteins are spotted onto a nitrocellulose sheet, and an aliquot of the covalent complex under test is added to each antibody spot. After suitable washing procedures, a positive reaction with one or other of the antisera is visualized by autoradiography of the labelled RNA moiety attached to the antibody via the ribosomal protein. Amounts of protein as low as 10 pg can readily be detected.
本文描述了一种在多组分系统研究中快速免疫鉴定蛋白质的方法。在这种情况下，系统是大肠杆菌核糖体，通过化学交联程序，待鉴定的核糖体蛋白与标记的核糖体RNA片段共价连接。针对各个核糖体蛋白产生的抗血清点样在硝酸纤维素膜上，将一份待测共价复合物加入到每个抗体点上。经过适当的洗涤程序后，通过对经由核糖体蛋白与抗体相连的标记RNA部分进行放射自显影，可观察到与一种或另一种抗血清的阳性反应。低至10 pg的蛋白量都能很容易地检测到。
