去垢剂法提取脂筏的量化  

The Quantification of Detergent Extraction Method of Lipid Rafts

作　　者：曲由[1] 聂坤[2] 韩鹦赢[3] 沈鹏[3] 尚磊[1] 陈璐[1] 于建春[2] 韩景献[2] 

机构地区：[1]天津中医药大学,天津300193 [2]天津中医药大学第一附属医院分子生物学研究所,天津300193 [3]天津市第一中心医院急症科,天津300193

出　　处：《现代生物医学进展》2012年第29期5752-5754,共3页Progress in Modern Biomedicine

基　　金：天津市应用基础与前沿技术研究计划重点项目(11JCZDJC20100);高等学校博士学科点专项科研基金(20101210110005);国家自然科学青年基金(30901927);天津市应用基础及前沿技术研究计划(09JCYBJC12600)

摘　　要：目的:将去垢剂法提取脂筏的操作方法量化。方法:依据脂质筏在4℃时不溶于去垢剂的特性提取脂筏,再用蔗糖密度梯度离心法将去垢剂不溶组分分离出来。用胆固醇吸光度及浮舰蛋白1(flotillin-1)作为脂质筏的特异性标记,验证提取物的特性。结果:在离心管5%蔗糖与30%蔗糖分界处看到一层连成片状乳黄色脂质物质,光散射法显示该提取物在620 nm处有最大吸光值,Western blot结果显示在相对分子质量48 kDa处可见条带。结论:提取物符合脂筏的多种特性,操作方法量化的去垢剂法是一种简单、稳定提取脂质筏的方法。ABSTRACT Objective： To quantify the method of using non-ionic detergent for extracting lipid rafts. Methods： Based on the characteristic of that lipid rafts is not soluable in non-ionic detergents at 4℃, lipid rafts could be extracted and then delaminated by sucrose gradient centrifugation. To validate the extract＇s characteristics, cholesterol absorbance and flotilin-1, a lipid rafts＇ specific markers, was detected. Results： A yellow lipide layer band located at the interface between 5 % and 30 % sucrose. It showed that there was a maximum absorbance value at 620 nm by light scattering and a band was visible at molecular mass 48 kDa site by Western blot. Conclusion： The extract corre- sponded with many characteristics of Iipid rafts. The quantified operation of detergent method is a simple and stable approach for lipid rafts extraction. Key words： Lipid rafts; Senescence Accelerated Mouse; Detergent

关 键 词：脂筏 SAM小鼠 去垢剂 

分 类 号：R284.2[医药卫生—中药学]