Going J J, Williams A R, Wyllie A H, Anderson T J, Piris J
Department of Pathology, University of Edinburgh, Scotland, U.K.
J Pathol. 1988 Jul;155(3):185-90. doi: 10.1002/path.1711550302.
Specific immunostaining of p21 ras protein by the well-characterized pan-ras antibody Y13-259 is achieved in paraffin sections of human and animal tissues fixed in periodate-lysine-paraformaldehyde-dichromate (PLPD). Intensity of staining is as good as in cryostat sections, with superior histological detail. Localization to plasma membrane is demonstrated in rodent cells genetically manipulated to express abundant p21 ras (the FHO5T1 cell line), both in preparations suspended in agar after culture in vitro and in those growing as tumour in vivo. Strong positive staining is observed in neoplasms of human breast and colon, tissues in which there is independent evidence of elevated ras gene expression. The superior morphology afforded by this technique allows clear characterization of p21 ras expression in small premalignant lesions for which other methods of detection of oncogene expression are not appropriate.
利用特性明确的泛ras抗体Y13 - 259，可在经高碘酸盐 - 赖氨酸 - 多聚甲醛 - 重铬酸盐（PLPD）固定的人和动物组织石蜡切片中实现p21 ras蛋白的特异性免疫染色。染色强度与冰冻切片相当，且组织学细节更优。在经基因操作以表达大量p21 ras的啮齿动物细胞（FHO5T1细胞系）中，无论是体外培养后悬浮于琼脂中的制剂，还是体内生长为肿瘤的制剂，均显示p21 ras定位于质膜。在人乳腺和结肠肿瘤中观察到强阳性染色，在这些组织中有独立证据表明ras基因表达升高。该技术提供的优越形态学特征使得能够清晰地鉴定小的癌前病变中p21 ras的表达情况，而其他检测癌基因表达的方法不适用于此类病变。
