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1、168中国实验血液学杂志JournalofExperimentalHematology2006;14(1):168-172文章编号(ArticleID):10092137(2006)01-0168-05论著几种双向凝胶电泳蛋白质检测方法的比较秦慧,刘霆,柳斌,宋鑫,黄欣,杨金亮,赵霞,魏于全四川大学华西医院血液科,人类疾病生物治疗国家重点实验室,成都610041摘要本研究对蛋白质染色法进行比较和分析,以求获得满足蛋白质组研究所需的适宜的蛋白质检测法。实验采用2D电泳分离急性早幼粒白血病细胞株NB4的全蛋白及1D电泳分离蛋白质分子量标准物,从蛋白质检测的灵敏性、
2、质谱兼容性、操作的简便性等角度,比较了传统考马斯亮蓝染色法、胶体考马斯亮蓝染色法、改良考马斯亮蓝染色法和银染法4种蛋白质着色法对凝胶分离蛋白质的检测影响,探讨了影响蛋白质染色与鉴定的影响因素。结果显示,银染检测灵敏度最高,但质谱鉴定相容性差,鉴定率近10%;改良考马斯亮蓝染色法检测灵敏度也高,可达4ng/spot,质谱鉴定率可接近55%。结论:改良考马斯亮蓝染色法的蛋白质检测灵敏度高,与质谱兼容好,能满足蛋白质组研究需要。关键词蛋白质组;双向凝胶电泳;蛋白质检测;考马斯亮蓝染色法;银染中图分类号Q503文献标识码AComparisonbetweenProteinDetec
3、tionMethodsforTwoDimensionalGelElectrophoresisQINHui,LIUTing,LIUBin,SONGXin,HUANGXin,YANGJinLiang,ZHAOXia,WEIYueQuanDepartmentofHematology,StateKeyLaboratoryofBiotherapyforHumanDiseases,WestChinaHospital,SichuanUniversity,Chengdu610041,ChinaAbstractTheaimofthisstudywastocompareandanalyzepr
4、oteinstaininginordertoselecttheoptimalstainingmethodforproteomicresearch.ProteinsfromacutepromyelocyticleukemiacelllineNB4andproteinmolecularweightmarkerwereseparatedrespectivelyby2Dor1DelectrophoresisanddetectedrespectivelybythetypicalCoomassiebrilliantblue,thecolloidalCoomassiebrilliantblue
5、,themodifiedCoomassiebrilliantblueandthesilverstainingprotocols.Theproteindetectionsensitivity,compatibilitywithmassspectrometry(MS)andfacilityofthefourstainingprotocolswerecompared.TheresultsindicatedthatthesilverstainingexhibitedthehighestsensitivityandMSshowedthelowestcompatibility10%ofprote
6、inidentificationrate.ThedetectionsensitivityofthemodifiedCoomassiebrilliantbluestainingwassuperiortothatofothertwoCoomassiebrilliantbluestainings,closetobutlowerthanthesilverstaining,howeverthecompatibilitywithMSwasbetter(proteinidentificationrateabout55%).Itisconcludedthattheproteindetectionse
7、nsitivityofthemodifiedCoomassiebrilliantbluestainingishigh,anditscompatibilitywithMSisbetter,thismodifiedCoomassiebrilliantbluestainingisanoptimalstainingmethodforproteomicresearch.Keywordsproteomics;twodimensionalgelelectropho
