刘友平,丁慧荣,肖斌,罗文炽,李洪

• 1:泸州医学院,泸州医学院,泸州医学院,泸州医学院,泸州医学院 (646000) ,(646000) ,(646000) ,(646000) ,(646000)

摘要：

目的　寻找一种更适合肝细胞蛋白质提取的细胞裂解液配方。方法　配制不同组成和不同浓度的细胞裂解液 ,分别对相同条件下培养的肝细胞进行裂解并进行蛋白定量及十二烷基磺酸钠 聚丙烯酰胺凝胶电泳 (SDS PAGE)佐证。结果　不同组成及不同浓度的细胞裂解液对肝细胞裂解作用存在显著差异。结论　去污剂用 4 % 3 [(3 胆酰胺丙基 ) 二乙胺 ] 丙磺酸 (CHAPS)的细胞裂解液对肝细胞裂解效果最好。

关键词： 肝细胞;蛋白;细胞裂解液

Abstract: Objective To search a suitable lysis buffer for hepatocytic protein extraction.Methods Prepare lysis buffers in different compositions and concentrations.Then hepatocytic proteins were collected through above buffers,hepatocytes incubated in the same condition were lysed separatelyandquantifiedwithBio-RadDC.SDS-PAGE was used to separate the hepatocytes proteins to testify the protein quantity.Results Lysis buffers of different compositions and concentrations had different effects on the cell lysis,with a statistical significance.Conclusion Lysis buffer containing 4% CHAPS (cholamidopropyl-dimethyl ammonio-propane sulfonate) detergent is the most suitable one for hepatocyte lysis.

Keywords: Hepatocytes; Egg white; Cell lysis buffer

基金项目: 国家自然科学基金资助项目 (70 0 710 4 0 );; 泸州医学院科研基金资助项目 (2 0 0 3 10 )

通讯作者:

Email:

参考文献：

• 1PenningtonSR ,DunnMJ,编著.蛋白质组学:从序列到功能.钱小红,译北京:科学出版社,2002.3851.
• 2WilmM ,ShevchenkoA ,HouthaeveT ,etal.Femtomolesequencingofproteinsfrompolyacrylamidegelsbynano electrospraymassspectrometry.Nature,1996,379(6564):466469.
• 3GonzalezJ,CornejoA ,SantosMR ,etal.Anovelproteinphos phatase2A (PP2A)isinvolvedinthetransformationofhumanpro tozoanparasiteTrypanosomacruzi.BiochemJ,2003,374(Pt3):647656.
• 4LiC ,HongY ,TanYX ,etal.Accuratequalitativeandquantitativeproteomicanalysisofclinicalhepatocellularcarcinomausinglasercapturemicrodissectioncoupledwithisotope codedaffinitytagandtwo dimensionalliquidchromatographymassspectrometry.MolCellProteomics,2004,3:399409.
• 5WangJ ,CheungAT ,KollsJK ,etal.Effectsofadenovirus mediatedliver selectiveoverexpressionofproteintyrosinephosphatase1boninsulinsensitivityinvivo.DiabetesObesMetab,2001,3(5):367380.
• 6HeileJM ,FongYL ,DomenicoR ,etal.EvaluationofhepatitisCvirusglycoproteinE2forvaccinedesign:anendoplasmicreticulum retainedrecombinantproteinissuperiortosecretedrecombinantproteinandDNA basedvaccinecandidates.Virol,2000,74(15):68856892.
• 7黄杰安,唐国都,丁虹,等.一种简单分离、传代培养成年小鼠肝细胞的方法.医学文选,2001,20(4):417418.