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Millipore/CC119 | Human Caspase 3, recombinant human active/CC119/25 units

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貨號: CC119

品牌: Millipore

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Description
CatalogueNumber	CC119
BrandFamily	Chemicon®
TradeName	Chemicon
Description	HumanCaspase3,recombinanthumanactive
Overview	Specificity:Caspase-3(alsoknownasCPP32,Yamaandapopain)isamemberoftheinterleukin-1betaconvertingenzyme(ICE)familyofcysteineproteases.Caspase-3existsincellsasaninactive32kDaproenzyme.Duringapoptosisprocaspase-3isprocessedataspartateresiduesbyself-proteolysisand/orcleavagebyupstreamcaspases,suchascaspase-6(Mch2),caspase-8(FLICE)granzymeB.Theprocessedformofcaspase-3consistsoflarge(17kDa)andsmall(12kDa)subunitswhichassociatetoformtheactiveenzyme.Theactivecaspase-3hasbeenshowninvolvingintheproteolysisofseveralimportantmolecules,suchaspoly(ADP-ribose)polymerase(PARP),thesterolregulatoryelementbindingproteins(SREBPs),focaladhesionkinase(FAK),andothers. Thepartiallypurifiedactiverecombinanthumancaspase-3proteinwithfull-lengthHis-6tagwasexpressedinE.coli.TheexpressedCaspase-3spontaneouslyundergoesautoprocessingtoyieldthesubunitscharacteristicofthenativeenzyme.Therecombinantcaspase-3preferentiallycleavescaspase-3substrates(e.g.,DEVD-AFCorDEVD-pNA).Incombinationwithcaspase-3activityassays,theactiverecombinantcaspase-3isveryusefulintheBIOLOGicalscreeningofcaspaseinhibitors.Therecombinantenzymecanalsobeusedasapositivecontrolincaspaseassaysorindeterminingthespecificityofcaspasesubstrates. 10-25%caspase-3proteinbygelanalysis,alsocontainsE.Coliproteins.Approximately,0.8-2μgtotalprotein/unitofactivity.GenBank#U13737.

ProductInformation
Presentation	Lyophilized.Reconstituteto1unit/μLwithPBScontaining15%glycerol. Partiallypurifiedrecombinantcaspase3withafulllengthHIS-6-tag.Approximately10-20%caspase3protein.TheremainderisE.Coliproteins.Approximately0.8-2mgtotalprotein/Uofactivity;E.Coliproteinshavenoreactivity.

StorageandShippingInformation
StorageConditions	Maintainunopenedandrehydratedproductat-70°Cinundiluted,freeze-thawprotected,tightlysealedaliquots.Avoidrepeatedfreeze/thawcycles.Productstablefor6monthsfromdateofpurchasedifhandledappropriately.

Applications
KeyApplications	ActivityAssay
ApplicationNotes	Westernblot:1-5μLofstockperlane.Proteinisfulllengthcaspase3withHis-Tag.Itisautoactivatedthusactivecaspasesubunitswillbeseen and17kDa+12kDa(largeandsmallsubunits)canbeseendependingupontheantibody.Smallsubunitisextremelydifficulttodetectinmostwesterns.Thesizeofinactiveenzimeis~32kDa. Activation:Notneeded.WhenexpressedinE.coli,caspase-3spontaneouslyundergoesautoprocessingtoyieldthesubunitscharacteristicoftheactiveenzyme.

Applications

KeyApplications

ActivityAssay

ApplicationNotes

Westernblot:1-5μLofstockperlane.Proteinisfulllengthcaspase3withHis-Tag.Itisautoactivatedthusactivecaspasesubunitswillbeseen

and17kDa+12kDa(largeandsmallsubunits)canbeseendependingupontheantibody.Smallsubunitisextremelydifficulttodetectinmostwesterns.Thesizeofinactiveenzimeis~32kDa.

Activation:Notneeded.WhenexpressedinE.coli,caspase-3spontaneouslyundergoesautoprocessingtoyieldthesubunitscharacteristicoftheactiveenzyme.

BiologicalInformation
Purity	Partiallypurifiedrecombinantcaspase3withafulllengthHIS-6-tag.Approximately10-20%caspase3protein.TheremainderisE.Coliproteins.Approximately0.8-2mgtotalprotein/Uofactivity;E.Coliproteinshavenoreactivity.
SpecificActivity	Oneunitoftherecombinantcaspase-3istheenzymeactivitythatcleaves1nmolofthecaspasesubstrateDEVD-pNA(pNA:pnitroanaline)perhourat37°Catsaturatedsubstrateconcentrations.
EntrezGeneNumber	NM_004346.3 NM_032991.2
EntrezGeneSummary	Thisgeneencodesaproteinwhichisamemberofthecysteine-asparticacidprotease(caspase)family.Sequentialactivationofcaspasesplaysacentralroleintheexecution-phaseofcellapoptosis.Caspasesexistasinactiveproenzymeswhichundergoproteolyticprocessingatconservedasparticresiduestoproducetwosubunits,largeandsmall,thatdimerizetoformtheactiveenzyme.Thisproteincleavesandactivatescaspases6,7and9,andtheproteinitselfisprocessedbycaspases8,9and10.Itisthepredominantcaspaseinvolvedinthecleavageofamyloid-beta4Aprecursorprotein,whichisassociatedwithneuronaldeathinAlzheimer"sdisease.Alternativesplicingofthisgeneresultsintwotranscriptvariantsthatencodethesameprotein.
GeneSymbol	CASP3 apopain CASP-3 procaspase3 CPP32B SCA-1 CPP32 Apopain CPP-32 Yama EC3.4.22.56[Contains:Caspase-3subunitp17 Caspase-3subunitp12].
UniProtNumber	P42574
UniProtSummary	FUNCTION:SwissProt:P42574#InvolvedintheactivationcascadeofcaspasesresponsIBLeforapoptosisexecution.Attheonsetofapoptosisitproteolyticallycleavespoly(ADP-ribose)polymerase(PARP)ata"216-Asp--Gly-217"bond.Cleavesandactivatessterolregulatoryelementbindingproteins(SREBPs)betweenthebasichelix-loop-helixleucinezipperdomainandthemembraneattachmentdomain.Cleavesandactivatescaspase-6,-7and-9.Involvedinthecleavageofhuntingtin. SIZE:277aminoacids;31608Da SUBUNIT:Heterotetramerthatconsistsoftwoanti-parallelarrangedheterodimers,eachoneformedbya17kDa(p17)anda12kDa(p12)subunit. SUBCELLULARLOCATION:Cytoplasm. TISSUESPECIFICITY:Highlyexpressedinlung,spleen,heart,liverandkidney.Moderatelevelsinbrainandskeletalmuscle,andlowintestis.Alsofoundinmanycelllines,highestexpressionincellsoftheimmunesystem. PTM:CleavagebygranzymeB,caspase-6,caspase-8andcaspase-10generatesthetwoactivesubunits.Additionalprocessingofthepropeptidesislikelyduetotheautocatalyticactivityoftheactivatedprotease.Activeheterodimersbetweenthesmallsubunitofcaspase-7proteaseandthelargesubunitofcaspase-3alsooccurandviceversa. SIMILARITY:SwissProt:P42574##BelongstothepeptidaseC14family.

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Millipore/CC119 | Human Caspase 3, recombinant human active/CC119/25 units

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