Henriksen A Z, Maeland J A
Department of Microbiology, Faculty of Medicine, University of Trondheim, Norway.
Acta Pathol Microbiol Immunol Scand B. 1987 Oct;95(5):315-21. doi: 10.1111/j.1699-0463.1987.tb03131.x.
The experimental conditions for antibody-binding by the 38.5 kD porin protein of an E. coli 055 strain in immunoblotting were investigated. A non-ionic detergent in the buffer which contained the primary antibody was required for antibody-binding by electroblots of the SDS-denatured protein. Immunoblotting, using antiserum absorbed with bacteria or the outer membrane (OM) of the E. coli 055 strain, showed results concordant with inaccessibility to antibodies of the 38.5 kD porin protein in its native configuration in the bacterial cells, but immunoreactivity when contained in the OM. OM from strains of different genera of the Enterobacteriaceae and antisera against these strains when used in immunoblot analyses showed that the E. coli 055 porin protein harboured antigenic determinants which are common to the various genera of the enteric bacilli. Cross-reactivity with non-enteric Gram-negative bacteria was not observed.
研究了大肠杆菌055菌株38.5 kD孔蛋白在免疫印迹中与抗体结合的实验条件。对于SDS变性蛋白的电印迹，在含有一抗的缓冲液中需要一种非离子去污剂才能实现抗体结合。使用经细菌或大肠杆菌055菌株外膜（OM）吸收的抗血清进行免疫印迹，结果表明，在细菌细胞中，天然构象的38.5 kD孔蛋白无法与抗体结合，但存在于OM中时具有免疫反应性。在免疫印迹分析中，使用肠杆菌科不同属菌株的OM以及针对这些菌株的抗血清，结果显示大肠杆菌055孔蛋白含有肠道杆菌各属共有的抗原决定簇。未观察到与非肠道革兰氏阴性菌的交叉反应。
