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Cytoskeleton/6xHis MICAL-1 Protein Redox-CH domains (2 x 50 micrograms)/1 x 1 mg/MIC01-XL
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4000-520-616
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MICAL(MIC01)isanintracellularflavoproteinmonoxygenase,conservedfrominsectstomammals,thatfunctionsasacatalystforoxidation-reduction(redox)reactions.Terman’sgroupshowedthatMICALinteractswithF-actinandusesNADPHasacofactortooxidizeactinatMet44andMet47.AspartoftheMOXtrue™productline,HumanMICAL1protein(MIC01)[accession#Q8TDZ2(MICA1_HUMAN)]hasbeenproducedandpurifiedfromabacterialexpressionsystem.TherecombinantproteinisN-terminally6xHistidinetaggedandiscomprisedoftheRedoxandCHdomains(aa:1-612)ofthefulllengthprotein.ValidationstudieshaveshownthatMIC01canoxidizeactinwhichwillreducesubtilisinAcleavageby>90%inlimitedproteolysisassays.MIC01hasalsobeenvalidatedfortheproductionofoxidizedactinwhichcanbeusedindownstreamapplicationssuchassedimentationassaysandactinpolymerizationassays.TolearnmoreaboutusingMICALasaresearchtoolseeourDatasheetTolearnmoreabouttheMICAL/MsrB/ActinphysiologicalredoxsystemseeourNewsletterEachlotofpurifiedproteinisqualitycontrolledtoprovidehighbatchtobatchconsistency,seeCOAdocuments.ValidatedApplicationsMICAL-1(MIC01)ProteinPurityDeterminationA10μgsampleofMICAL-1proteinwassepa-ratedbyelectrophoresisina4-20%Tris-GlycinegelandstainedwithCoomassieBlue.ProteinquantitationwasperformedusingthePrecisionRedTMProteinAssayReagent(Cat.#ADV02).Mark12standardmolecularweightMarkersarefromInvitrogen.SubtilisinAssayonMICAL-1TreatedActinActin(Cat#AKL99)wasdilutedto0.1mg/ml(2.3mM)andleftuntreated,ortreatedwithNADPH(400mM),MIC01(0.076mM),oracombinationofboth(seemethod).2mgofeachsamplewasthenleftuntreated,ortreatedwithsubtilisin(1:200w/w)for15min.SampleswerethenseparatedbySDS-PAGEandvisualizedwithCoomassiestaining.ClickhereforadetailedmethodActinSedimentation+/-MICAL-1TreatmentActin(Cat#AKL99)wasdilutedto0.2mg/ml(4.6mM)or0.8mg/ml(18.4mM)(seemethod).Sampleswerethenincubatedwith2xpolymerizationbufferor2xpolymerizationbuffersupplementedwithMIC01+NADPHfor1hatroomtemperatire.Sampleswerespuninanultracentrifugeat100,000gfor1.5h.SampleswerethenseparatedbySDS-PAGEandvisualizedwithCoomassiestaining.ClickhereforadetailedmethodActinPolymerization+/-MICAL-1Treatment.Pyrenelabeled-actin(Cat.#AP05)wasdilutedto0.1mg/ml(2.3mM)(seemethod).Sampleswerethenincubatedwith2xpolymerizationbuffersupplementedwithnothing(labeledControl),NADPHalone(labeled+NADPH),MIC01alone(labeled+MIC01),orthecombinationofboth(labeled+Both).Uponactinpolymerizationfluorescencewasdetectedwithaspectrophotometer(seemethodbelow).A.U.=arbitraryunitsClickhereforadetailedmethodFormoreinformationcontact:signalseeker@Cytoskeleton.comAssociatedProducts:MOXtrue™6xHisMsrB2Protein(Cat.#MB201)MOXtrue™MICAL-oxidizedActin(Cat.#MXA95)MOXtrue™MICAL-oxidizedPyreneLabeledActin(Cat.#MXAP95)RabbitSkeletalMuscleActin(Cat.#AKL95)PyreneLabeledRabbitSkeletalMuscleActin(Cat.#AP05)
Cytoskeleton，Inc.成立于1993年。自成立以来，我们一直在不断扩大产品范围。
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