使用抗异硫氰酸荧光素单克隆抗体通过半抗原介导鉴定细胞膜抗原。

Hapten-mediated identification of cell membrane antigens using an anti-FITC monoclonal antibody.

作者信息

Balogh P, Szekeres G, Németh P

机构信息

Immunological and Biotechnological Laboratory, University Medical School of Pécs, Hungary.

出版信息

J Immunol Methods. 1994 Feb 28;169(1):35-40. doi: 10.1016/0022-1759(94)90122-8.

DOI:10.1016/0022-1759(94)90122-8

PMID:7510754

Abstract

A monoclonal anti-FITC antibody (F4/1) was produced and demonstrated to be specific for both the free and protein-conjugated (either soluble or cell-bound) form of fluorescein, or carboxyfluorescein. When mouse thymocytes were labelled with a novel fluorescein derivative 5(6)-carboxyfluorescein succinimidyl ester (CFl-NSE), the incorporation of fluorescein was predominantly membrane-bound as demonstrated immunohistochemically. The coupling of CFl-NSE to cells displays a random distribution pattern as shown by immunoblotting of cell extracts prepared by detergent solubilization of CFl-NSE-labelled thymocytes. In addition, the Thy-1.2 antigen immunoprecipitated from a CFl-NSE-labelled thymocyte lysate with a rat monoclonal antibody (Mab) could be detected using the anti-FITC Mab. The molecular weight of the immunoprecipitated material could be estimated immediately by reference to the FITC-labelled molecular weight markers electrophoresed simultaneously.

摘要

制备了一种单克隆抗异硫氰酸荧光素抗体（F4/1），并证明其对游离形式和蛋白质结合形式（可溶性或细胞结合形式）的荧光素或羧基荧光素具有特异性。当用新型荧光素衍生物5(6)-羧基荧光素琥珀酰亚胺酯（CFl-NSE）标记小鼠胸腺细胞时，免疫组织化学显示荧光素的掺入主要是膜结合的。如通过去污剂溶解CFl-NSE标记的胸腺细胞制备的细胞提取物的免疫印迹所示，CFl-NSE与细胞的偶联呈现随机分布模式。此外，使用抗异硫氰酸荧光素单克隆抗体可以检测到用大鼠单克隆抗体（Mab）从CFl-NSE标记的胸腺细胞裂解物中免疫沉淀的Thy-1.2抗原。通过同时电泳的异硫氰酸荧光素标记的分子量标志物可以立即估计免疫沉淀物质的分子量。