碱性成纤维细胞生长因子（bFGF）在源自大鼠邓宁前列腺癌的转移性细胞系（AT-3）中的定位：一种特异性单克隆抗体的应用

Localization of basic fibroblast growth factor (bFGF) in a metastatic cell line (AT-3) established from the Dunning prostatic carcinoma of rat: application of a specific monoclonal antibody.

作者信息

Nishikawa K, Yoshitake Y, Minemura M, Yamada K, Matuo Y

机构信息

Department of Biochemistry, Kanazawa Medical University, Ishikawa, Japan.

出版信息

Adv Exp Med Biol. 1992;324:131-9. doi: 10.1007/978-1-4615-3398-6_13.

DOI:10.1007/978-1-4615-3398-6_13

PMID:1283495

Abstract

Localization of basic fibroblast growth factor (bFGF) in a metastatic cell line, AT-3, established from the Dunning prostatic carcinoma of rat was determined by two immunological techniques using a specific monoclonal antibody against bFGF. Concentration of bFGF in cell extract was measured by sandwich radioimmunoassay (RIA) with heparin-Sepharose and 125I-labeled monoclonal antibody. bFGF concentration in the extract of AT-3 cells increased with increasing concentration of NaCl in extraction buffer. Localization of bFGF in AT-3 cells was determined by counting radioactivity of 125I-labeled monoclonal antibody binding to AT-3 cells before or after increasing permeability of the cells. The binding increased significantly by this treatment, indicating that bFGF within the cells was detected.

摘要

利用针对碱性成纤维细胞生长因子（bFGF）的特异性单克隆抗体，通过两种免疫技术确定了从大鼠邓宁前列腺癌建立的转移性细胞系AT-3中bFGF的定位。用肝素-琼脂糖和125I标记的单克隆抗体通过夹心放射免疫测定法（RIA）测量细胞提取物中bFGF的浓度。AT-3细胞提取物中bFGF的浓度随提取缓冲液中NaCl浓度的增加而增加。通过计数增加细胞通透性前后与AT-3细胞结合的125I标记单克隆抗体的放射性来确定AT-3细胞中bFGF的定位。通过这种处理，结合显著增加，表明检测到了细胞内的bFGF。