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通过用合成肽免疫制备兔抗小鼠CD8抗原复合物抗血清及其特性鉴定

Production and characterization of a rabbit antiserum to the mouse CD8 antigenic complex by immunization with a synthetic peptide.

作者信息

Brunati S, Corradin G, Bron C

出版信息

J Immunol Methods. 1987 Jan 26;96(1):97-105. doi: 10.1016/0022-1759(87)90372-3.

Abstract

A rabbit antiserum to the mouse CD8 antigen (Lyt-2/3) was obtained through the use of a synthetic peptide corresponding to the N-terminal segment of the 37 kDa subunit of the CD8 molecular complex. This anti-peptide antiserum detected specifically the membrane antigen from detergent extracts of surface-labeled mouse thymocytes. The efficiency of the immunoprecipitation increased significantly upon treatment of the cell lysates with 0.1% SDS at 100 degrees C before immunopurification. Finally both the 37 kDa and the 32 kDa polypeptides expressing the Lyt-2 antigen were revealed by immunoblotting.

摘要

通过使用与CD8分子复合物37 kDa亚基N端片段相对应的合成肽,获得了针对小鼠CD8抗原(Lyt-2/3)的兔抗血清。这种抗肽抗血清能特异性地检测来自表面标记小鼠胸腺细胞去污剂提取物中的膜抗原。在免疫纯化前,将细胞裂解物在100℃用0.1% SDS处理后,免疫沉淀效率显著提高。最后,通过免疫印迹法揭示了表达Lyt-2抗原的37 kDa和32 kDa多肽。

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