Fernández E, Kopchick J J
Department of Zoological and Biomedical Sciences, Ohio University, Athens 45701.
Anal Biochem. 1990 Dec;191(2):268-71. doi: 10.1016/0003-2697(90)90218-x.
Blotting techniques have been extensively used, not only analytically for protein identification, but also preparatively to isolate and purify specific proteins from a large variety of cellular extracts and biological fluids. The process involves the separation of proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transfer of proteins to nitrocellulose membranes, and immunostaining to identify proteins which often are at very low concentrations. Because of the quantitative interactions of the proteins with specific antibodies, we have coupled the immunoblot procedure with photographic and densitometric methods for the quantitative determination of bovine growth hormone (bGH). In this way, the method is suitable for bGH detection and quantitation for a small number of samples by use of a single Western blot analysis. The sensitivity of this method permits determinations of bGH to 0.5 ng. The method uses a comparative procedure in which purified bovine growth hormone is used as a standard.
印迹技术不仅广泛用于蛋白质鉴定分析，还用于从各种细胞提取物和生物体液中分离和纯化特定蛋白质。该过程包括通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离蛋白质，将蛋白质转移到硝酸纤维素膜上，以及通过免疫染色鉴定通常浓度很低的蛋白质。由于蛋白质与特异性抗体的定量相互作用，我们将免疫印迹程序与摄影和光密度测定方法相结合，用于定量测定牛生长激素（bGH）。通过这种方式，该方法适用于通过单次蛋白质印迹分析对少量样品进行bGH检测和定量。该方法的灵敏度可检测至0.5 ng的bGH。该方法采用一种比较程序，其中使用纯化的牛生长激素作为标准品。
