Deo S S, Fule R P, Saoji A M
Department of Microbiology, Dr. V.M. Medical College, Solapur, India.
Indian J Exp Biol. 1990 Dec;28(12):1184-6.
Prestaining of human serum proteins with a new reactive dye Drimarene Brilliant Blue (DBB), was standardized employing 940 separations and examining 30 variables. Under the critical condition, the serum and the soluble dye (0.1 g/100 ml in working Tris-glycine buffer, pH, 8.3), was mixed in equal proportion, conjugate warmed at 40 degrees C for 2 hr and a 30 microliter of the sample electrophoresed by disc electrophoresis. The method when compared with prestaining by Remazol Brilliant Blue (RBB) and postelectrophoretic staining by Amido Black (AB) in 50 normal sera, revealed that the discs stained with DBB were intense and well defined and appeared in 2 hr on a sparkingly clear gel. Quality of resolution was better than RBB and AB. Protein bands eluted from the DBB prestained gels retained their immunoreactivity. The dye-protein complex of albumin and transferrin produced high-titre monospecific antisera in rabbits.
采用一种新型活性染料直接耐晒翠蓝（DBB）对人血清蛋白进行预染色，并通过940次分离和检测30个变量进行了标准化。在临界条件下，将血清与可溶性染料（在工作Tris-甘氨酸缓冲液中，pH值为8.3，0.1 g/100 ml）等比例混合，共轭物在40℃下温育2小时，取30微升样品进行圆盘电泳。与在50份正常血清中用雷马素亮蓝（RBB）预染色和用氨基黑（AB）进行电泳后染色相比，该方法显示用DBB染色的圆盘条带颜色深且清晰，在2小时内出现在晶莹剔透的凝胶上。分辨率质量优于RBB和AB。从DBB预染色凝胶上洗脱的蛋白条带保留了它们的免疫反应性。白蛋白和转铁蛋白的染料-蛋白质复合物在兔体内产生了高滴度的单特异性抗血清。
