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Millipore/APT171 | Caspase 8 Colorimetric Activity Assay Kit, IETD/APT171/100 assays

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貨號: APT171

品牌: Millipore

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Description
CatalogueNumber	APT171
BrandFamily	Chemicon®
TradeName	Chemicon
Description	Caspase8ColorimetricActivityAssayKit,IETD
Overview	ActivationofICE-familyproteases/caspasesinitiatesapoptosisinmammaliancells.Chemicon"sCaspase-8ColorimetricActivityAssayKitsprovideasimpleandconvenientmeansforassayingtheactivityofcaspasesthatrecognizethesequenceIETD.Theassayisbasedonspectophotometricdetectionofthechromophorep-nitroaniline(pNA)aftercleavagefromthelabeledsubstrateIETD-pNA.ThefreepNAcanbequantifiedusingaspectrophotometeroramicrotiterplatereaderat405nm.ComparisonoftheabsorbanceofpNAfromanapoptoticsamplewithanuninducedcontrolallowsdeterminationofthefoldincreaseincaspase-8activity.
AlternateNames	FLICE
MaterialsRequiredbutNotDelivered	·Microcentrifugeand1.5mLMicrocentrifugetubes ·37°CWaterbathorIncubator ·SpectrophotometerorMicroplateReader ·96wellMicrotiterPlate

ProductInformation
Components	5XCellLysisBuffer(PartNo.90065):5mL 5XAssayBuffer(PartNo.90066):10mL Caspase-8Substrate(Ac-IETD-pNA)(PartNo.90082):1mLof2.5mg/mLsolution Caspase-8Inhibitor(Ac-IETD-CHO)(PartNo.90086):50μLof100μM(0.05mg/mL)inDMSO pNAStandard(PartNo.90085):250μLof10mMinDMSO
Detectionmethod	Chromogenic

StorageandShippingInformation
StorageConditions	Storekitmaterialsat-20°Cuptotheirexpirationdate. SpecialPrecautions: ·Afterthawingreagents,useimmediatelyoraliquotandfreezeat-20oCforlongerstorage.Avoidrepeatedfreeze/thawcycles. ·TheCaspase-8substrateandpNAstandardareespeciallylightsensitive.Maintainthesereagentsinamberorcoveredcontainers.

Applications
Application	Caspase-8ColorimetricActivityAssayKitsprovideasimple&convenientmeansforassayingtheactivityofcaspasesthatrecognizethesequenceIETD.
KeyApplications	ActivityAssay

BIOLOGicalInformation
SpeciesReactivity	Mammals
EntrezGeneNumber	NM_001080124.1 NM_001080125.1 NM_001228.4 NM_033355.3 NM_033356.3 NM_033358.3
EntrezGeneSummary	Thisgeneencodesamemberofthecysteine-asparticacidprotease(caspase)family.Sequentialactivationofcaspasesplaysacentralroleintheexecution-phaseofcellapoptosis.Caspasesexistasinactiveproenzymescomposedofaprodomain,alargeproteasesubunit,andasmallproteasesubunit.Activationofcaspasesrequiresproteolyticprocessingatconservedinternalasparticresiduestogenerateaheterodimericenzymeconsistingofthelargeandsmallsubunits.ThisproteinisinvolvedintheprogrammedcelldeathinducedbyFasandvariousapoptoticstimuli.TheN-terminalFADD-likedeatheffectordomainofthisproteinsuggeststhatitmayinteractwithFas-interactingproteinFADD.ThisproteinwasdetectedintheinsolublefractionoftheaffectedbrainregionfromHuntingtondiseasepatientsbutnotinthosefromnormalcontrols,whichimplicatedtheroleinneurodegenerativediseases.Manyalternativelysplicedtranscriptvariantsencodingdifferentisoformshavebeendescribed,althoughnotallvariantshavehadtheirfull-lengthsequencesdetermined.
GeneSymbol	CASP8 MCH5 MGC78473 CASP-8 MACH ALPS2B procaspase-8 FLICE CAP4 EC3.4.22.61[Contains:Caspase-8subunitp18 Caspase-8subunitp10].
UniProtNumber	Q14790
UniProtSummary	FUNCTION:SwissProt:Q14790#MostupstreamproteaseoftheactivationcascadeofcaspasesresponsIBLefortheTNFRSF6/FASmediatedandTNFRSF1Ainducedcelldeath.BindingtotheadaptermoleculeFADDrecruitsittoeitherreceptor.Theresultingaggregatecalleddeath-inducingsignalingcomplex(DISC)performsCASP8proteolyticactivation.TheactivedimericenzymeisthenliberatedfromtheDISCandfreetoactivatedownstreamapoptoticproteases.ProteolyticfragmentsoftheN-terminalpropeptide(termedCAP3,CAP5andCAP6)arelikelyretainedintheDISC.CleavesandactivatesCASP3,CASP4,CASP6,CASP7,CASP9andCASP10.MayparticipateintheGZMBapoptoticpathways.CleavesADPRT.Hydrolyzesthesmall-moleculesubstrate,Ac-Asp-Glu-Val-Asp--AMC.LikelytargetforthecowpoxvirusCRMAdeathinhibitoryprotein.Isoforms5,6,7and8lackthecatalyticsiteandmayinterferewiththepro-apoptoticactivityofthecomplex. SIZE:479aminoacids;55391Da SUBUNIT:Heterotetramerthatconsistsoftwoanti-parallelarrangedheterodimers,eachoneformedbya18kDa(p18)anda10kDa(p10)subunit.InteractswithFADD,CFLARandPEA15.Isoform9interactsattheendoplasmicreticulumwithacomplexcontainingBCAP31,BAP29,BCL2and/orBCL2L1. SUBCELLULARLOCATION:Cytoplasm. TISSUESPECIFICITY:Isoforms1,5and7areexpressedinawidevarietyoftissues.Highestexpressioninperipheralbloodleukocytes,spleen,thymus,andliver.Barelydetectableinbrain,testis,andskeletalmuscle.DOMAIN:SwissProt:Q14790Isoform9containsaN-terminalextensionthatisrequiredforinteractionwiththeBCAP31complex. PTM:Generationofthesubunitsrequiresassociationwiththedeath-inducingsignalingcomplex(DISC),whereasadditionalprocessingislikelyduetotheautocatalyticactivityoftheactivatedprotease.GZMBandCASP10canbeinvolvedintheseprocessingevents.&PhosphorylateduponDNAdamage,probablybyATMorATR. DISEASE:SwissProt:Q14790#DefectsinCASP8arethecauseofcaspase-8deficiency(CASP8D)[MIM:607271].CASP8Disadisorderresemblingautoimmunelymphoproliferativesyndrome(ALPS).Itischaracterizedbylymphadenopathy,splenomegaly,anddefectiveCD95-inducedapoptosisofperipheralbloodlymphocytes(PBLs).ItleadstodefectsinactivationofT-lymphocytes,B-lymphocytes,andnaturalkillercellsleADIngtoimmunodeficiencycharacterizedbyrecurrentsinopulmonaryandherpessimplexvirusinfectionsandpoorresponsestoimmunization. SIMILARITY:BelongstothepeptidaseC14family.&Contains2DED(deatheffector)domains.

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Millipore/APT171 | Caspase 8 Colorimetric Activity Assay Kit, IETD/APT171/100 assays

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