Giardina S L, Evans S W, Gandino L, Robey F A, Bonvini E, Longo D L, Varesio L
Anal Biochem. 1987 Feb 15;161(1):109-16. doi: 10.1016/0003-2697(87)90659-2.
A hybridoma producing a monoclonal antibody (MoAB) recognizing both the cellular and viral forms of fos has been generated by somatic cell hybridization techniques from spleen cells of mice immunized with a synthetic peptide corresponding to amino acids 128-152, a consensus region, of both the v-fos and c-fos oncogene products. Three proteins with molecular weights of 55,000, 44,000, and 42,000 were detected by immunoblotting. While MoAB 2G9C3 failed to immunoprecipitate fos from Finkel-Biskis-Jenkins murine osteosarcoma-virus-infected fibroblasts, both the 55,000 v-fos protein and the 39,000 cellular protein were coprecipitated using polyvalent rabbit antibodies to the same peptide. Whereas no cell surface membrane expression of fos was detected, after membrane permeabilization by a brief exposure to lysolecithin it was possible to specifically detect internal fos by immunofluorescence flow cytometry. Immunohistochemical staining of FBJ virus-infected cells revealed intense, nuclear staining.
通过体细胞杂交技术，利用对应于v-fos和c-fos癌基因产物共有区域（氨基酸128 - 152）的合成肽免疫小鼠的脾细胞，产生了一种能识别fos细胞形式和病毒形式的单克隆抗体（MoAB）的杂交瘤。通过免疫印迹法检测到三种分子量分别为55,000、44,000和42,000的蛋白质。虽然MoAB 2G9C3无法从芬克尔-比斯克斯-詹金斯鼠骨肉瘤病毒感染的成纤维细胞中免疫沉淀fos，但使用针对同一肽的多价兔抗体可共沉淀55,000的v-fos蛋白和39,000的细胞蛋白。虽然未检测到fos的细胞表面膜表达，但通过短暂暴露于溶血卵磷脂使细胞膜通透后，可通过免疫荧光流式细胞术特异性检测细胞内的fos。对FBJ病毒感染细胞的免疫组织化学染色显示出强烈的核染色。
