Shalitin C, Kenoshi E, Bartal A H
Department of Biology, Technion-Israel Institute of Technology, Haifa.
Cancer Invest. 1988;6(6):687-94. doi: 10.3109/07357908809078036.
Rabbit polyclonal antibodies (pAb) were raised against a yeast ras-related protein YP20 and shown to be immunoreactive with human normal as well as altered Ha-ras and Ki-ras p21 gene products using immunoblotting and immunoprecipitation techniques. The p21 protein revealed by anti-YP20 antibodies comigrates with p21 protein detected by anti-p21 monoclonal antibody (Cetus Diagnostics). These pAbs were tested against a panel of human acetone-fixed tumor cell lines and malignant effusions and nonfixed fresh-frozen tissue sections obtained from cancer patients by the indirect immunofluorescence assay (IFA). Twelve of sixteen (75%) sarcoma and carcinoma cells lines and one fibroblast cell line were stained by the anti-YP20 pAb. The binding occurred most commonly in the cytoplasm. Six of eight fresh-frozen colon and breast cancer tissue sections were immunostained and normal sections from these organs or skin showed only low level of binding to the pAbs. Four of five malignant effusions were distinctively immunostained. These antibodies are suggested to serve as additional probes for assessing the expression of ras gene-related proteins in human malignancy.
用酵母ras相关蛋白YP20制备兔多克隆抗体（pAb），并采用免疫印迹和免疫沉淀技术证明其与人正常以及发生改变的Ha-ras和Ki-ras p21基因产物具有免疫反应性。抗YP20抗体所识别的p21蛋白与抗p21单克隆抗体（Cetus诊断公司）检测到的p21蛋白迁移率相同。通过间接免疫荧光分析（IFA），用这些pAb检测了一组人丙酮固定的肿瘤细胞系、恶性积液以及取自癌症患者的未固定新鲜冷冻组织切片。16个肉瘤和癌细胞系中的12个（75%）以及1个成纤维细胞系被抗YP20 pAb染色。结合最常见于细胞质中。8个新鲜冷冻的结肠癌和乳腺癌组织切片中有6个被免疫染色，而这些器官或皮肤的正常切片与pAb的结合水平很低。5个恶性积液中有4个被明显免疫染色。这些抗体被认为可作为评估人恶性肿瘤中ras基因相关蛋白表达的额外探针。
