CHEMILUMINESCENT ENHANCED CD47 DETECTION ON WESTERN BLOTTING

CD47 is a leucocyte antigen of wide tissue distribution including plat elets and erythrocytes. It has been found that anti-CD47 antibodies ma y cause platelet tion of platelet glycoproteins including CD47. Conven tional Western blotting techniques using horseradish peroxidase-conjug ated antibody and 3,3'-diaminobenzidine (DAB) substrate resulted in un satisfactory results with CD47 due to low sensitivity. When chemilumin escent detection with horseradish peroxidase-conjugated antibody was u sed in conjunction with Western blotting, detection of the CD47 molecu le was greatly enhanced. A protein of 47-55 kDa immunoblotted from pla telets 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 u seful and sensitive alternative for the study of human platelet antige ns.