蛋白质相关实验技术操作指南

精品文档1、Westernblotting:WesternblottingusingECLreagentusedby：LaboratoryofP.J.Hansen,Dept.ofAnimalSciences,UniversityofFloridaKamps'sWesternBlottingProtocolusedby：SeftonLab,SalkInstituteDryTransfer 干法蛋白转膜usedby：ThePreussLab,TheDivisionofBiologicalSciences,TheUniversityofChicagoWesternBlotofTBPfromTBP-GSTbacteriausedby：StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSFWesternBlot/Anti-P-CREBusedby：StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSFWesternblottingprotocolfor1C2,3B5H10,and4C8Antibodiesusedby：StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSFProtocolforanti-HAantibodyWesternBlottingusedby：StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSFProbingandStrippingofWesternBlotusedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstituteWesternBlottingProtocolusedby：MirmiraLaboratoryattheUniversityofVirginiaWesternBlottingandImmunostainingusedby：ChazinLab,CenterforStructuralBiology,VanderbiltUniversityWesternBlotsusedby：LabofDr.MarkBartonFrank,OklahomaMedicalResearchFoundationWesternBlotting-Antibodiesusedby：TheMinionLab,CollegeofVeterinaryMedicineatIowaStateUniversityWesternBlottingusedby：Cepko/TabinLab,HarvardUniversityCarbonateSolutionForWesternBlotusedby：Dr.DEKoshland,CarnegieInstitutionofWashingtonWesternBlotwithBSAusedby：Dr.DEKoshland,CarnegieInstitutionofWashingtonMETHODforWesternBlotsusedby：Dr.DEKoshland,CarnegieInstitutionofWashingtonWesternBlotsusedby：MichaelKoelle'sLab,DepartmentofMolecularBiophysicsandBiochemistryYaleUniversity2、蛋白分离/提取/浓缩。1欢迎下载精品文档LysisBuffersusedby：StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSFYeastProteinIsolation"Yaffe-Schatz"usedby：AmbergLab,UpstateMedicalUniversityGuanidineHydrochloridePurificationofProteinsFromSDSPAGEusedby：GottschlingLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstituteYeastproteinprepforSDSPAGEandwestern(rapid)usedby：GottschlingLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstituteYeastproteinprepforSDSPAGEandwestern(glassbead)usedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstitutePurificationofTFIIICfromyeastusedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstitutePurificationofTFIIAfromE.coliusedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstituteTCAproteinprecipitationusedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstitutePurificationofMLCK(MyosinLightChainKinase)fromRabbitSkeletalMuscle 肌球蛋白轻链激酶usedby：StullLab,UniversityofTexas,SouthwesternMedicalCenterPurificationofCalmodulinfromBovineBrainusedby：StullLab,UniversityofTexas,SouthwesternMedicalCenterMyosinLightChainPreparationfromSkeletalandCardiacMuscleusedby：StullLab,UniversityofTexas,SouthwesternMedicalCenterPurifyingProteinfromInclusionBodiesusedby：ChazinLab,CenterforStructuralBiology,VanderbiltUniversityProtocolforProteinExtractionfromPlantusedby：TheUniversityofNebraska-LincolnProteinCoreFacility(PCF)TaqPolymerasePurificationusedby：Cepko/TabinLab,HarvardUniversityYeastProteinPrep(NoBoil)usedby：Dr.DEKoshland,CarnegieInstitutionofWashingtonYeastProteinPrep(Boil)usedby：Dr.DEKoshland,CarnegieInstitutionofWashingtonGSTFusionProteinPrepusedby：VesicleTrafficking,StanfordUniversity3、蛋白质检测分析：AMIDOBLACKSTAINusedby：HancockLaboratoryMethods,DepartmentofMicrobiologyandImmunology,UniversityofBritishColumbia,BritishColumbia,Canada。2欢迎下载精品文档JamesHardwick'sangiotensinassayprotocolusedby：SeftonLab,SalkInstitutePhosphoaminoacidanalysis:MarkKamps'smethodusedby：SeftonLab,SalkInstituteULTIMATEHIS-UBASSAYFORusedby：theTanseyLabatColdSpringHarborLaboratoryULTIMATEHIS-UBASSAYFORYEASTusedby：theTanseyLabatColdSpringHarborLaboratoryProtocolforBeta-HexosaminidaseRBL-2H3SecretionAssayusedby：meyerlab,StanfordUniversityMeasurementofMyosinRegulatoryLightChain(RLC)Phosphorylationusedby：StullLab,UniversityofTexas,SouthwesternMedicalCenterBicinchoninicAcid(BCA)ProteinAssayusedby：DavidR.Caprette,RiceUniversityBiuretProteinAssayusedby：DavidR.Caprette,RiceUniversityBradfordproteinassayusedby：DavidR.Caprette,RiceUniversityHartree-LowryandModifiedLowryProteinAssaysusedby：DavidR.Caprette,RiceUniversityColorimetricAssaysusedby：DavidR.Caprette,RiceUniversityKdmeasurement,fitting,calculation,andsimulationusedby：ChazinLab,CenterforStructuralBiology,VanderbiltUniversity4、蛋白标记修饰DSPCrosslinkingusedby：PeterNovickLab,DepartmentofCellBiologyYaleUniversitySchoolofMedicineLabelingGc-GlobulinwithCarboxyfluoresceinSuccinimidylEsterusedby：Yu-LiWangLab,UniversityofmassachusettsMedicalSchool(UMASS)LabelingTubulinwithTetramethylrhodamineSuccinimidylEsterusedby：Yu-LiWangLab,UniversityofmassachusettsMedicalSchool(UMASS)LabelingGizzardVinculinwithTetramethylrhodamineIodoacetamideusedby：Yu-LiWangLab,UniversityofmassachusettsMedicalSchool(UMASS)LabelingGizzardAlpha-actininwithTetramethylrhodamineIodoacetamideusedby：Yu-LiWangLab,UniversityofmassachusettsMedicalSchool(UMASS)LabelingGizzardMyosinwithTetramethylrhodamineIodoacetamide(primarily17kDMLC)usedby：Yu-LiWangLab,UniversityofmassachusettsMedicalSchool(UMASS)LabelingGizzardMyosinwithTetramethyorhodamineIodoacetamide(MHCand17kDMLC)。3欢迎下载精品文档usedby：Yu-LiWangLab,UniversityofmassachusettsMedicalSchool(UMASS)LabelingMuscleActinwithN-(1-pyrene)Iodoacetamideusedby：Yu-LiWangLab,UniversityofmassachusettsMedicalSchool(UMASS)LabelingMuscleActinwithTetramethylrhodamineIodoacetamideusedby：Yu-LiWangLab,UniversityofmassachusettsMedicalSchool(UMASS)LabelingMuscleActinwithCarboxyfluoresceinSuccinimidyleEsterusedby：Yu-LiWangLab,UniversityofmassachusettsMedicalSchool(UMASS)LabelingMuscleActinwith5-Iodoacetamidofluoresceinusedby：Yu-LiWangLab,UniversityofmassachusettsMedicalSchool(UMASS)ultimatemammaliancellpulse-chaseusedby：theTanseyLabatColdSpringHarborLaboratoryultimateyeastpulse-chaseusedby：theTanseyLabatColdSpringHarborLaboratoryGluteraldehydeConjugationofOligopeptidestoProteinsusedby：LabofDr.MarkBartonFrank,OklahomaMedicalResearchFoundationFluoresceinlabelingofproteinsusedby：DavidChambers,SalkinstituteBiosyntheticlabelingusedby：SeftonLab,SalkInstitute,SanDiego,California5、免疫沉淀(IP/CoIP/ChIP)Immunoprecipitationbuffersandprotocolsusedby：StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSFCellLysis/Western/IPusedby：StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSFAmplificationandLabellingwithCydyesusedby：YaleGenomeAnalysisCenter,YaleUniversityChromatinImmunoprecipitationfromMammalianCellExtractsusedby：YaleGenomeAnalysisCenter,YaleUniversityChromatinImmunoprecipitationfromYeastWholeCellExtractsusedby：YaleGenomeAnalysisCenter,YaleUniversityImmunoprecipitationusedby：HancockLaboratoryMethods.DepartmentofMicrobiologyandImmunology,UniversityofBritishColumbia,BritishColumbia,CanadaUltimateyeastdenaturingIPusedby：theTanseyLabatColdSpringHarborLaboratoryULTIMATEFREEZE-THAWLYSISFORusedby：theTanseyLabatColdSpringHarborLaboratoryUltimatemammaliannondenaturingIP。4欢迎下载精品文档usedby：theTanseyLabatColdSpringHarborLaboratoryUltimatemammaliandenaturingIPusedby：theTanseyLabatColdSpringHarborLaboratoryUltimateyeastChIPassayusedby：theTanseyLabatColdSpringHarborLaboratoryChromatinImmunoprecipitationAssayandPCRusedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstituteYeastChromatinImmunoprecipitation(ChIP)usedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstituteMirmiraLabChIPProtocolusedby：MirmiraLaboratoryattheUniversityofVirginiaChIPAssayProtocolusedby：MirmiraLaboratoryattheUniversityofVirginiaImmunoprecipitationandImmuneComplexkinaseassayusedby：SeftonLab,SalkInstituteGeneralPrinciplesofImmunoprecipitationusedby：SeftonLab,SalkInstitute6、融合蛋白表达及纯化UseofB-PERlysisreagenttopurifyrecombinantproteinstaggedwithHis6ormaltosebindingproteinusedby：SmithLab,DivisionofBiologicalSciences,UniversityofMissouriBacterialExpressionofKinesinMotorsusedby：LizGreeneandSteveHenikoffPurificationofrecombinantsBRFM166Lusedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstituteProteinExpressionandPurificationofRPA70-ABusedby：ChazinLab,CenterforStructuralBiology,VanderbiltUniversityCo-ExpressionandpurificationofhumanDNAprimasep49-p58subunits引发酶p49-p58亚基共表达纯化方法usedby：ChazinLab,CenterforStructuralBiology,VanderbiltUniversityProteinExpressionandPurificationofMBP-TantigenMBP-T抗原表达纯化usedby：ChazinLab,CenterforStructuralBiology,VanderbiltUniversityPerspectivesonBaculovirusExpressionSystems表达usedby：LabofDr.MarkBartonFrank,OklahomaMedicalResearchFoundationExpression&PurificationofHisTaggedProteinsinE.coliusedby：Cepko/TabinLab,HarvardUniversity。5欢迎下载精品文档7、相互作用分析Amplifyingalargephage-displaylibrarywithoutlosingdiversityusedby：SmithLab,DivisionofBiologicalSciences,UniversityofMissouriProteinInteractionAnalysisUsinganIASYSBiosensorusedby：KittoLab,TheUniversityofTexasatAustinYeastTwo-HybridScreenwithLibraryandBaitusedby：MirmiraLaboratoryattheUniversityofVirginia8、蛋白电泳/双向电泳SDSusedby：StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSFGelDrying干胶usedby：StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSFNuPAGEGelElectrophoresis蛋白电泳usedby：KittoLab,TheUniversityofTexasatAustinIsoelectricFocussingofMembraneProteinsbySlabGelMethodusedby：HancockLaboratoryMethods,DepartmentofMicrobiologyandImmunology,UniversityofBritishColumbia,BritishColumbia,CanadaSilverStainingSDSPAGEGelsusedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstituteE.colitotalproteinforSDSPAGEusedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstituteComassieBlueProteingelstainusedby：HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstituteCoomassie&SilverStainingofPolyacrylamideGelsusedby：LabofDr.MarkBartonFrank,OklahomaMedicalResearchFoundation2-DPolyacrylamideGelElectrophoresisusedby：MolecularProfilingInitiative,NationalCancerInstitute(NCI)SilverStainingofSDSGelsusedby：TheMinionLab,CollegeofVeterinaryMedicineatIowaStateUniversitySDSGelsusedby：TheMinionLab,CollegeofVeterinaryMedicineatIowaStateUniversityCuCl2StainingofSDSGelsusedby：Cepko/TabinLab,HarvardUniversityElectroelutionofProteinsFromSDSGelsusedby：Drs.CCepkoandCTabin,HarvardUniversitySilverStainofProteinGels。6欢迎下载精品文档usedby：Dr.DEKoshland,CarnegieInstitutionofWashingtonSDS（BIO-RADusedby：Dr.DEKoshland,CarnegieInstitutionofWashingtonSDSusedby：MichaelKoelle'sLab,DepartmentofMolecularBiophysicsandBiochemistryYaleUniversity9、质谱分析Polycrystallinethinfilms 质谱样品准备usedby：ProwlLab,RockefellerUniversitySeededfilms(Vorm-Roepstorff)usedby：ProwlLab,RockefellerUniversityDrieddropletusedby：ProwlLab,RockefellerUniversityIn-gelTrypticDigestforProteinIDbyMassSpectrometryusedby：MitshisonLab,DepartmentofSystemsBiology,HarvardMedicalSchool10、crystallizationPolycrystallinethinfilmsusedby：PROWL,RockefellerUniversitySlowcrystallizationusedby：PROWL,RockefellerUniversityCrystallizationofKinesinFamilyMotorProteins 驱动蛋白家族的结晶条件和晶体参数usedby：LizGreeneandSteveHenikoffCrystallizationTrialsusedby：LizGreeneandSteveHenikoff11、invitroproteinsynthesisProteinSynthesesinCellFreeSystems。7欢迎下载精品文档usedby：TheLaboratoryofWilliamHeidcampatGustavusAdolphusUniversity12、蛋白纯化/层析6xHis-taggedproteinpurificationusingQiagenNi-NTAColumnunderNativeConditionsusedby：Sugdenlab,McArdleLaboratoryforCancerResearch,UniversityofWisconsin-MadisonMedicalSchoolPurificationofdnEBNA-1/SoftfromE.coliBL21LysSusedby：Sugdenlab,McArdleLaboratoryforCancerResearch,UniversityofWisconsin-MadisonMedicalSchoolPurificationofGSTfusionproteinsinE.coliGST 融合蛋白纯化（方法四）筛选表达株usedby：Sugdenlab,McArdleLaboratoryforCancerResearch,UniversityofWisconsin-MadisonMedicalSchoolPurificationofGSTfusionproteinsinE.coliGST 融合蛋白纯化，方法三，纯化小量蛋白usedby：Sugdenlab,McArdleLaboratoryforCancerResearch,UniversityofWisconsin-MadisonMedicalSchoolPurificationofGSTfusionproteinsinE.coliGST 融合蛋白纯化，方法二usedby：Sugdenlab,McArdleLaboratoryforCancerResearch,UniversityofWisc