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

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

目的 :提取 Mc Ab 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 :Mc Ab SZ- 39识别的蛋白呈一条区带 ,分子量为 6 0 k D。结论 :该 6 0 k D的蛋白是 Mc Ab SZ-

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. [