人血清P21蛋白的Western印迹测定  

Western blot detection of P21 protein in human serum

作　　者：贾定武[1] 吴逸明[1] 陈琛[1] 高亚胜 

机构地区：[1]河南医科大学劳动卫生学教研室,洛阳华美生物工程公司

出　　处：《河南医科大学学报》1995年第1期17-20,共4页Journal of Henan Medical University

基　　金：研究生课题

摘　　要：建立了Ｗｅｓｔｅｒｎ印迹法测定人血清中ｒａｓ癌基因蛋白的方法，检测限值为３ｍｇ／Ｌ血清。丽春红Ｓ染色后裁掉非靶蛋白部分硝酸纤维素膜，可大量节约一抗的用量；微波炉促进免疫结合，加速显色过程；用国产碱性磷酸酶ＡＢＣ试剂，可降低成本；实验结果可用５６０ｎｍ波长的色谱扫描或血清系列稀释法进行定量，并已用于职业肿瘤高危人群的筛检。Western blot method for detecting ras gene protein in human serum was establlshed,and as little as 3 mg of P21 protein in one liter of human serum could be determined. The excision of non-target-protein part of the electroblotted nitrocellulose membrane after Ponceau S staining minimized the consumption of the primary antibody. Microwave oven promoted immunobinding and shortened the whole detection period. The application of homemade ABC-CIAP kit for the visualization of bindings made the method possible to be popularized domestically. The result evaluation can be reached with dual wavelength chromato scanning at the wavelength of 560 nm or with serial dilution of serum. The method has been applied to assess occupational cancer risk.

关 键 词：蛋白 印迹 WESTERN印迹 血清 P21蛋白 

分 类 号：R446.11[医药卫生—诊断学]