Ramírez P, Bonilla J A, Moreno E, León P
J Immunol Methods. 1983 Aug 12;62(1):15-22. doi: 10.1016/0022-1759(83)90105-9.
Peroxidase-bound protein A and lectins in combination with precipitable substrates were used to detect viral proteins and glycoproteins on 'Western blots'. The sensitivity of various methods was compared, including radioiodinated protein A, enzyme immunosorbent assay and peroxidase staining with DAP. The sensitivity of the peroxidase reaction, its rapidity and low cost compared with other methods make this a useful choice for developing protein bands transferred to nitrocellulose paper. In addition, a procedure was devised to clarify and fix nitrocellulose sheets for direct quantification of the precipitated substrate by densitometric analysis.
将过氧化物酶结合蛋白A和凝集素与可沉淀底物相结合,用于在“蛋白质免疫印迹法”中检测病毒蛋白和糖蛋白。比较了包括放射性碘化蛋白A、酶联免疫吸附测定和用二氨基联苯胺进行过氧化物酶染色等各种方法的灵敏度。与其他方法相比,过氧化物酶反应的灵敏度、速度和低成本使其成为显影转移到硝酸纤维素纸上的蛋白条带的一个有用选择。此外,还设计了一种程序来澄清和固定硝酸纤维素膜,以便通过光密度分析直接定量沉淀的底物。