血清蛋白乙酸纤维素薄膜电泳方法的优化  

Optimization of Cellulose Acetate Film Electrophoresis Method for Serum Proteins

作　　者：闫巧梅[1] 李斌[1] 张学明[1] 孔凡青[1] 高英辰 丁海麦[1] Yan Qiaomei;Li Bin;Zhang Xueming;Kong Fanqing;Gao Yingchen;Ding Haimai(Baotou Medical College,Baotou 014040,China)

机构地区：[1]包头医学院,内蒙古包头014040

出　　处：《黑龙江科学》2024年第2期79-81,86,共4页Heilongjiang Science

基　　金：包头医学院科创融合项目(BYJJ-KCRH202305);包头医学院课程思政专项教学改革项目(2022KCSZ-18);包头医学院“问学计划”项目(2021BYWWJ-YB-15);内蒙古自治区教育科学研究“十四五”规划课题(NGJG H2022200)。

摘　　要：乙酸纤维素薄膜电泳是生物化学与分子生物学实验常规项目之一,临床上广泛应用于血清蛋白、糖蛋白及核酸等生物大分子的分离与检测。该实验受环境和人为因素影响较多,初次操作往往难以取得理想的实验结果,更换乙酸纤维素薄膜厂家后,原来的实验条件分离血清蛋白存在蛋白条带聚集、分不开现象。缓冲液离子浓度和成分是影响乙酸纤维素电泳的最关键因素,采用pH 8.6,离子强度0.025 M/L、0.075 M/L的巴比妥缓冲液;pH 8.6,离子强度0.05 M/L、0.08 M/L的硼酸缓冲液,用4种缓冲液对新膜进行筛选,并对电泳时长及上样进行优化。结果表明,用pH 8.6、离子强度为0.08 M/L硼酸缓冲液,电压100 V,电泳时长50 min,分离兔血清蛋白实验结果比较理想,能满足实验教学及临床检验要求。Cellulose acetate film electrophoresis is one of the routine projects in biochemistry and molecular biology experiments,and it is widely used for clinical separation and detection of biological macromolecules,such as serum proteins and glycoproteins.This experiment is influenced by many factors,and the initial operation often makes it difficult to achieve ideal experimental results.After changing the manufacturer of cellulose acetate film,the original experimental conditions for separating serum proteins show protein band aggregation,which cannot be separated.The ion concentration and composition of buffer are the most important influencing factors of cellulose acetate electrophoresis.Barbital buffer with pH 8.6,ionic strength of 0.025 M/L and 0.075 M/L is used in this experiment.PH 8.6,ionic strength of 0.05 M/L and 0.08M/L boric acid buffer,four kinds of buffer are used to screen the new membrane,and the electrophoresis duration and sample loading are optimized.The result shows that the separating rabbit serum protein of pH 8.6,ionic strength of 0.08 M/L boric acid buffer,voltage of 100 V,electrophoresis time of 50 min is ideal,which can meet the requirements of experiment teaching and clinical experiments.

关 键 词：乙酸纤维素薄膜电泳 硼酸缓冲液 生物化学与分子生物学实验教学 

分 类 号：R446.1[医药卫生—诊断学]