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AntibodiesOnline/Protein Kinase C, gamma (PRKCG) (Active) Protein/ABIN411969/5 μg188bio精品生物—专注于实验室精品爆款的电商平台 - 蚂蚁淘旗下精选188款生物医学科研用品
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AntibodiesOnline/Protein Kinase C, gamma (PRKCG) (Active) Protein/ABIN411969/5 μg
ProteinName
ProteinKinaseC,gamma(PRKCG)
  • PRKCG
  • PKC-gamma
  • PKCC
  • PKCG
  • SCA14
  • PKCgamma
  • Pkcc
  • Prkcc
  • PKC
  • PKCI
  • Prkc
  • RATPKCI
  • proteinkinaseC,gamma
  • PRKCG
  • Prkcg
Origin
Human
13Human
3Rat(Rattus)
1Mouse(Murine)
Source
BaculovirusinfectedInsectCells
5InsectCells
3Escherichiacoli(E.coli)
1BaculovirusinfectedInsectCells
1Baculovirus/Sf9
1HEK-293Cells
1Sf-9cells
1Wheatgerm
1Yeast
ProteinType
Recombinant
BIOLOGicalActivity
Active
Application
FunctionalStudies(Func),SDS-PAGE(SDS),WesternBlotting(WB)
Options
Bulkdiscount
Supplier
SupplierProductNo.
Purpose>90%PureactiveRecombinantPKCgamma.
CharacteristicsProteinSource:Baculovirus(Sf9insectcells)
PuritySDS-PAGE:≥80 %
BackgroundPKCγisamemberoftheproteinkinaseC(PKC)familyofserine-andthreonine-specificproteinkinasesthatcanphosphorylateawidevarietyofproteintargetsknowntobeinvolvedindiversecellularsignalingpathways.Inthebrain,PKCistranslocatedtocellmembranesduringischemiaandisrapidlyremovedordegradedduringthesecondotherwiselethalischemicinsultinpreconditionedbrains.ThissuggestthatischemicpreconditioningenhancesdownregulationofcellsignalingmediatedbyPKCandmaytherebyprovideneuroprotection(1).
AlternateNames:PKC,proteinkinaseC,gammaproteinkinaseC
MolecularWeight105.0kDa
GeneID5582
NCBIAccessionNM_002739
ResearchAreaNeurology
PathwaysWNTSignaling,EGFRSignalingPathway,NeurotrophinSignalingPathway,ThyroidHormoneSynthesis,MyometrialRelaxationandContraction,G-proteinmediatedEvents,PositiveRegulationofResponsetoDNADamageStimulus,InteractionofEGFRwithphospholipaseC-gamma,ThromboxaneA2ReceptorSignaling
ApplicationNotesOptimalworkingdilutionshouldbedeterminedbytheinvestigator.
Comment

ActivitySpecification:ThespecificactivityofPKCγwasdeterminedtobe512nmol/min/mgasperactivityassayprotocol.

ProtocolNote:thesearesuggestedworkingdilutionsanditisrecommendedthattheresearcherperformaserialdilutionofActivePKCgammaforoptimalresults).[32P]-ATPAssayCocktailPrepare250μM[32P]-ATPAssayCocktailinadesignatedrADIoactiveworkingareabyaddingthefollowingcomponents:150µLof10mMATPStockSolution,100µL[32P]-ATP(0mCi/100µL),5.70mlofKinaseAssayBuffer.Store0mlaliquotsat-20°C.KinaseDilutionBuffer,pH7.2KinaseAssayBufferIdilutedata1:4ratio(5Xdilution)withdistilledH2O.10mMATPStockSolutionPrepareATPstocksolutionbydissolving50mgofATPin10mlofKinaseAssayBuffer.Store200µLaliquotsat-20°C.KinaseAssayBufferI,pH7.2Buffercomponents:20mMMOPS,12.0mMbeta-glycerol-phosphate,20mMMgC12,0mMEGTA,0mMEDTA.Add0.20mMDTTtoKinaseAssayBufferpriortouse.SubstratePKCsyntheticpeptidesubstrate(ERMRPRKRQGSVRRRV)dilutedindistilledH2Otoafinalconcentrationof0mg/mL.AssayProtocolStep1.Thaw[32P]-ATPAssayCocktailinshieldedcontainerinadesignatedradioactiveworkingarea.Step2.ThawtheActivePKCgammaamma,KinaseAssayBuffer,SubstrateandEnzymeDilutionBufferonice.Step3.Inapre-cooledmicrofugetube,addthefollowingreactioncomponentsbringingtheinitialreactionvolumeupto20µL:Component1.10µLofdilutedActivePKCgamma.Sonicatelipidfor1minutepriortouse.
4.Setuptheblankcontrolasoutlinedinstep3,excludingtheadditionofthesubstrate.ReplacethesubstratewithanequalvolumeofdistilledH2O.
5.Initiatethereactionbytheadditionof0µL[32P]-ATPAssayCocktailbringingthefinalvolumeupto20µLandincubatethemixtureinawaterbathat30°Cfor15minutes.
6.Afterthe15minuteincubationperiod,terminatethereactionbyspotting20µLofthereactionmixtureontoindividualpre-cutstripsofphosphocelluloseP81paper.
7.Airdrythepre-cutP81stripandsequentiallywashina1%phosphoricacidsolution(dilute10mlofphosphoricacidandmakea1LsolutionwithdistilledH2O)withconstantgentlestirring.Itisrecommendedthatthestripsbewashedatotalof3intervalsforapproximately10minuteseach.
8.CounttheradioactivityontheP81paperinthepresenceofscintillationfluidinascintillationcounter.
9.Determinethecorrectedcpmbyremovingtheblankcontrolvalue(seeStep4)foreachsampleandcalculatethekinasespecificactivityasoutlinedbelow.
RestrictionsForResearchUseonly
FormatLiquid
Concentration0.1mg/mL
BufferRecombinantproteinstoredin50 mMTris-HCl, pH7.5,150 mMNaCl,0.25 mMDTT,0.1 mMEGTA,0.1 mMEDTA,0.1 mMPMSF,25 %glycerol.
PreservativeDithiothreitol(DTT)
PrecautionofUseThisproductcontainsdithiothreitol(DTT):aPOISONOUSANDHAZARDOUSSUBSTANCEwhichshouldbehandledbytrainedstaffonly.
HandlingAdviceCentrifugethevialpriortoopening.
Storage-80°C
StorageComment-80°C
ExpiryDate12months
SupplierImages
Western Blotting (WB) image for Protein Kinase C, gamma (PRKCG) (Active) Protein (ABIN411969)ProteinKinaseC,gamma(PRKCG)(Active)protein

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