Lalani E N, Golding M, Hudson M, Chieffi G, Stamp G, Anilkumar T V, Sarraf C, Alison M R
Department of Histopathology, Royal Postgraduate Medical School, London, U.K.
J Pathol. 1995 Nov;177(3):323-8. doi: 10.1002/path.1711770316.
Polyacrylamide gel electrophoresis (PAGE) of proteins with subsequent western blotting has become a routine technique for the analysis of proteins from both cultured cells and fresh whole tissue. We have developed a method to extract proteins from methacarn-fixed tissue which renders them suitable for SDS-PAGE and western blotting. With a panel of antibodies to specific intermediate filaments, transforming growth factor-alpha (TGF-alpha), and albumin, immunohistochemistry was performed in parallel with western blotting on sections cut from methacarn-fixed samples of normal rat liver and liver from rats treated under a regime which induces oval cell proliferation. Immunohistochemistry enabled the determination of changes in tissue distribution and abundance of the target proteins, which was mirrored by the corresponding western blot data. This technique can be especially effective when used in conjunction with immunohistochemistry. Tissue samples are easy to prepare, avoiding the precautions which need to be taken when handling fresh tissue (Abstract: J Pathol 1994; 173S: No. 41).
蛋白质的聚丙烯酰胺凝胶电泳（PAGE）及随后的蛋白质免疫印迹法已成为分析培养细胞和新鲜全组织中蛋白质的常规技术。我们开发了一种从甲醇-冰醋酸固定组织中提取蛋白质的方法，使其适用于SDS-PAGE和蛋白质免疫印迹法。使用一组针对特定中间丝、转化生长因子-α（TGF-α）和白蛋白的抗体，对从正常大鼠肝脏以及在诱导卵圆细胞增殖的方案下处理的大鼠肝脏的甲醇-冰醋酸固定样本切片进行蛋白质免疫印迹的同时进行免疫组织化学。免疫组织化学能够确定靶蛋白的组织分布和丰度变化，相应的蛋白质免疫印迹数据也反映了这一点。该技术与免疫组织化学结合使用时可能特别有效。组织样本易于制备，避免了处理新鲜组织时需要采取的预防措施（摘要：《病理学杂志》1994年；173S：第41期）
