-
公开(公告)号：KR1020130114407A
公开(公告)日：2013-10-18
申请号：KR1020120036738
申请日：2012-04-09
Applicant: 충남대학교산학협력단
CPC classification number: C07H1/06 , C12Y305/01052 , G01N27/62 , G01N2400/10 , H01J49/26
Abstract: PURPOSE: A method for selectively isolating a cell membrane for detecting sugar chains contained in the cell membrane by mass spectroscopy is provided to effectively isolate the cell membrane and to enhance sensitivity and accuracy in sugar chain analysis. CONSTITUTION: A method for selectively isolating a cell membrane for detecting sugar chains contained in the cell membrane by mass spectroscopy comprises the steps of: adding a stabilizing buffer solution to cells, which contains sodium hydrate and/or potassium hydroxide, a protease inhibitor, and sugar and does not contain a surfactant and a chelating agent; treating the container containing the buffer solution and the cells on the ice by ultrasonic waves and destructing the cells; centrifuging the cell lysate and removing a nuclear fraction and residues; adding the buffer solution to the supernatant; ultracentrifuging the supernatant mixture and removing a cytoplasmic fraction; resuspending the precipitate in a Na2CO3 solution (pH 11) and destroying microsome; ultracentrifuging the secondary mixture twice and removing the supernatant containing the cytoplasmic fraction; and isolating a cell membrane fraction.
Abstract translation: 目的：提供用于通过质谱法选择性分离用于检测细胞膜中所含糖链的细胞膜的方法，以有效地分离细胞膜并提高糖链分析的灵敏度和准确性。 构成：用于通过质谱法选择性分离用于检测细胞膜中所含糖链的细胞膜的方法包括以下步骤：将稳定缓冲溶液加入含有水合钠和/或氢氧化钾的蛋白酶抑制剂和 糖，不含表面活性剂和螯合剂; 通过超声波处理含有缓冲溶液的容器和冰上的细胞并破坏细胞; 离心细胞裂解物并除去核心部分和残留物; 将缓冲溶液加入上清液; 超离心上清混合物并除去细胞质级分; 将沉淀物重悬于Na 2 CO 3溶液（pH11）中并破坏微粒体; 将二次混合物超速离心两次并除去含有细胞质级分的上清液; 并分离细胞膜级分。
-
Search Results
2
records
(took 0.015 seconds)
