摘要
目的优化点样材料、缓冲液浓度和点样量,提高血浆蛋白醋酸纤维薄膜电泳(简称电泳)效果。方法用滤纸条、火车票、订书钉点等量血浆于稀释4倍(0.25×)Na2HPO4-KH2PO4缓冲液(简称缓冲液)中电泳,比较电泳效果,探索最佳点样材料;用滤纸条点等量血浆,比较不同稀释倍数的缓冲液电泳效果,探索最佳缓冲液浓度;用滤纸条点不同量的血浆于最佳缓冲液中电泳,比较电泳效果,探索最佳点样量。结果三种点样材料依次可见6、6、5条带;滤纸条点样的薄膜条带最清晰,火车票次之,订书钉最差。1×、0.5×、0.33×、0.25×、0.2×的缓冲液电泳后分别得到6、6、6、6、5条带,0.5×缓冲液所得的6条带最清晰。点样0.5μL的薄膜只见5条带;点样1、2、3、4μL的薄膜均能见6条带,以点样2μL的薄膜6条带最为清晰。结论用滤纸条点样2μL血浆、用稀释两倍的缓冲液电泳能获得最佳电泳效果。
Objective The sample-applying material,buffer concentration and sample size were optimized to improve the effect of plasma protein acetate cellulose membrane electrophoresis(abbreviated as ACME).Methods The same amount of plasma applied with filter paper strip,train ticket and staple pointers repectively was seperated by ACME under the 0.25×Na2HPO4-KH2PO4buffer solution(abbreviated as BS),to compare the electrophoresis effect and explore the best sample-applying material.The same amount of plasma applied with filter paper strips was separated in different concentration of BS,the electrophoresis results were compared and the optimal BS concentrations was determined.The different amounts of plasma applied with filter paper strips were separated in the optimal BS to compare the electrophoresis effect and explore the optimal sample size.Results In this study,when applied by filter paper strips,the plasma was separated as six bands which were the most clear and least trailing.When BS diluted twice,plasma was separated as six most clear protein bands.When the plasma was two microliters,six clear bands were visible,and the fifth band was the clearest.Conclusion The effect of plasma electrophoresis was greatly improved by applying filter paper strip,diluting BS two times and applying two microliter of plasma.
作者
姚蕾
李雨勤
吴永亮
黎威巍
金科华
YAO Lei;LI Yu-qin;JIN Ke-hua(Biochemistry Department,School of Basic Medicine,Hubei University of Science and Technology,Xianning Hubei 437100,China)
出处
《湖北科技学院学报（医学版）》
2020年第1期11-13,F0002,共4页
Journal of Hubei University of Science and Technology(Medical Sciences)
基金
湖北科技学院科研探索课题(ZJ0467)。
关键词
点样材料
缓冲液浓度
点样量
Sample-appling material
Buffer concentration
Sample size
作者简介
通讯作者:金科华,E-mail:striveapple@126.com。
