单克隆抗体SZ—39识别的人脑胶质瘤细胞SHG—44膜蛋白的提取

目的：提取ＭｃＡｂ ＳＺ－３９识别的人脑胶质瘤细胞ＳＨＧ－４４膜抗原蛋白，为其进一步研究作准备。方法：分别以两组去污剂（１）２％Ｔｒｉｔｏｎ Ｘ－１１４；（２）１％ＳＤＳ、１％去氧胆酸钠、１％ＮＰ－４０、１ｍｍｏｌ／Ｌ ＥＤＴＡ裂解ＳＨＧ－４４细胞，提取细胞蛋白，再以ＳＤＳ－ＰＡＧＥ法分离蛋白，并以Ｗｅｓｔｅｒｎ－Ｂｌｏｔ法分析鉴定。结果：ＭｃＡｂ ＳＺ－３９识别的蛋白呈一条区带，分子量为６０ｋＤ

EXTRACTION OF THE MEMBRANE PROTEIN OF HUMAN GLIOMA CELL SHG-44 IDENTIFIED BY McAb SZ-39

Objective: Extract the membrane protein of human glioma cell SHG-44 identified by McAb-39 to prepare for the further study of such as assay of amino acid sequence. Methods: Glioma cells were split with two groups of detergent (1)2% Triton X-114;(2)1% SDS, 1% Deoxycholic acid Na salt, 1% NP-40, 1mmol/L EDTA to extract cell proteins respectively, then SDS-PAGE was adopted to separate the proteins and Western-Blot was adopted to identify the specific protein. Results: Only one positive band appeared after SDS-PAGE and Western-Blot of the two kinds of protein extract respectively. The two positive bands were of a same molecular weight, 60kD. Conclusion: The 60kD protein is the antigen identified by McAb SZ-39 or short peptide contains the specific epitope.