蛋白质相关实验技术操作指南.doc

蛋白质相关实验技术操作指南.doc

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1、Westernblotting:

WesternblottingusingECLreagent

usedby：

LaboratoryofP.J.Hansen,Dept.ofAnimalSciences,UniversityofFlorida

Kamps'sWesternBlottingProtocol

usedby：

SeftonLab,SalkInstitute

DryTransfer干法蛋白转膜

usedby：

ThePreussLab,TheDivisionofBiologicalSciences,TheUniversityofChicago

WesternBlotofTBPfromTBP-GSTbacteria

usedby：

StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSF

WesternBlot/Anti-P-CREB

usedby：

StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSF

Westernblottingprotocolfor1C2,3B5H10,and4C8Antibodies

usedby：

StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSF

Protocolforanti-HAantibodyWesternBlotting

usedby：

StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSF

ProbingandStrippingofWesternBlot

usedby：

HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstitute

WesternBlottingProtocol

usedby：

MirmiraLaboratoryattheUniversityofVirginia

WesternBlottingandImmunostaining

usedby：

ChazinLab,CenterforStructuralBiology,VanderbiltUniversity

WesternBlots

usedby：

LabofDr.MarkBartonFrank,OklahomaMedicalResearchFoundation

WesternBlotting-Antibodies

usedby：

TheMinionLab,CollegeofVeterinaryMedicineatIowaStateUniversity

WesternBlotting

usedby：

Cepko/TabinLab,HarvardUniversity

CarbonateSolutionForWesternBlot

usedby：

Dr.DEKoshland,CarnegieInstitutionofWashington

WesternBlotwithBSA

usedby：

Dr.DEKoshland,CarnegieInstitutionofWashington

METHODforWesternBlots

usedby：

Dr.DEKoshland,CarnegieInstitutionofWashington

WesternBlots

usedby：

MichaelKoelle'sLab,DepartmentofMolecularBiophysicsandBiochemistry

YaleUniversity

2、蛋白分离/提取/浓缩

LysisBuffers

usedby：

StevenFinkbeiner,DepartmentsofNeurologyandPhysiology,UCSF

YeastProteinIsolation"Yaffe-Schatz"

usedby：

AmbergLab,UpstateMedicalUniversity

GuanidineHydrochloridePurificationofProteinsFromSDSPAGE

usedby：

GottschlingLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstitute

YeastproteinprepforSDSPAGEandwestern（rapid）

usedby：

GottschlingLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstitute

YeastproteinprepforSDSPAGEandwestern（glassbead）

usedby：

HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstitute

PurificationofTFIIICfromyeast

usedby：

HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstitute

PurificationofTFIIAfromE.coli

usedby：

HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstitute

TCAproteinprecipitation

usedby：

HahnLab，TheFredHutchinsonCancerResearchCenterandHowardHughesMedicalInstitute

PurificationofMLCK（MyosinLightChainKinase）fromRabbitSkeletalMuscle肌球蛋白轻链激酶

usedby：

StullLab,UniversityofTexas,SouthwesternMedicalCenter

PurificationofCalmodulinfromBovineBrain

usedby：

StullLab,UniversityofTexas,SouthwesternMedicalCenter

MyosinLightChainPreparationfromSkeletalandCardiacMuscle

usedby：

StullLab,UniversityofTexas,SouthwesternMedicalCenter

PurifyingProteinfromInclusionBodies

usedby：

ChazinLab,CenterforStructuralBiology,VanderbiltUniversity

ProtocolforProteinExtractionfromPlant

usedby：

TheUniversityofNebraska-LincolnProteinCoreFacility（PCF）

TaqPolymerasePurification

usedby：

Cepko/TabinLab,HarvardUniversity

YeastProteinPrep（NoBoil）

usedby：

Dr.DEKoshland,CarnegieInstitutionofWashington

YeastProteinPrep（Boil）

usedby：

Dr.DEKoshland,CarnegieInstitutionofWashington

GSTFusionProteinPrep

usedby：

VesicleTrafficking,StanfordUniversity

3、蛋白质检测分析：

AMIDOBLACKSTAIN

usedby：

HancockLaboratoryMethods,DepartmentofMicrobiologyandImmunology,

UniversityofBritishColumbia,BritishColumbia,Canada

JamesHardwick'sangiotensinassayprotocol

usedby：

SeftonLab,SalkInstitute

Phosphoaminoacidanalysis:

MarkKamps'smethod

usedby：

SeftonLab,SalkInstitute

ULTIMATEHIS-UBASSAYFOR

usedby：

theTanseyLabatColdSpringHarborLaboratory

ULTIMATEHIS-UBASSAYFORYEAST

usedby：

theTanseyLabatColdSpringHarborLaboratory

ProtocolforBeta-HexosaminidaseRBL-2H3SecretionAssay

usedby：

meyerlab,StanfordUniversity

MeasurementofMyosinRegulatoryLightChain（RLC）Phosphorylation

usedby：

StullLab,UniversityofTexas,SouthwesternMedicalCenter

BicinchoninicAcid（BCA）ProteinAssay

usedby：

DavidR.Caprette,RiceUniversity

BiuretProteinAssay

usedby：

DavidR.Caprette,RiceUniversity

Bradfordproteinassay

usedby：

DavidR.Caprette,RiceUniversity

Hartree-LowryandModifiedLowryProteinAssays

usedby：

DavidR.Caprette,RiceUniversity

ColorimetricAssays

usedby：

DavidR.Caprette,RiceUniversity

Kdmeasurement,fitting,calculation,andsimulation

usedby：

Chazi