Yuan F F, Mihrshahi S, Fletcher A
NSW Red Cross Blood Transfusion Service, Sydney, Australia.
Electrophoresis. 1996 Jan;17(1):219-20. doi: 10.1002/elps.1150170136.
CD47 is a leucocyte antigen of wide tissue distribution including platelets and erythrocytes. It has been found that anti-CD47 antibodies may cause platelet activation. Sensitive techniques are therefore needed for biochemical investigation of platelet glycoproteins including CD47. Conventional Western blotting techniques using horseradish peroxidase-conjugated antibody and 3,3'-diaminobenzidine (DAB) substrate resulted in unsatisfactory results with CD47 due to low sensitivity. When chemiluminescent detection with horseradish peroxidase-conjugated antibody was used in conjunction with Western blotting, detection of the CD47 molecule was greatly enhanced. A protein of 47--55 kDa was immunoblotted from platelets by anti-CD47 while the mAb immunoblotted a broader band (45--60 kDa) from erythrocyte membranes. Thus, there may be different forms of CD47 present on platelets and erythrocytes. The results indicate that Western blotting with the chemiluminescent detection technique is a useful and sensitive alternative for the study of human platelet antigens.
CD47是一种白细胞抗原，广泛分布于包括血小板和红细胞在内的多种组织中。已发现抗CD47抗体可能会导致血小板活化。因此，对于包括CD47在内的血小板糖蛋白进行生化研究需要灵敏的技术。使用辣根过氧化物酶偶联抗体和3,3'-二氨基联苯胺（DAB）底物的传统蛋白质印迹技术，由于灵敏度低，对CD47的检测结果不尽人意。当将辣根过氧化物酶偶联抗体的化学发光检测与蛋白质印迹结合使用时，CD47分子的检测得到了极大的增强。抗CD47从血小板中免疫印迹出一条47-55 kDa的蛋白质条带，而单克隆抗体从红细胞膜中免疫印迹出一条更宽的条带（45-60 kDa）。因此，血小板和红细胞上可能存在不同形式的CD47。结果表明，采用化学发光检测技术的蛋白质印迹法是研究人类血小板抗原的一种有用且灵敏的替代方法。
