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Millipore/MAB4078 | Anti-PCNA Antibody, clone 131-11912/MAB4078/100 µL

價格: ￥3576.00 市場價: 5960.00

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貨號: MAB4078

品牌: Millipore

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Description
CatalogueNumber	MAB4078
BrandFamily	Chemicon®
TradeName	Chemicon
Description	Anti-PCNAAntibody,clone131-11912
AlternateNames	ProliferatingCellNuclearAntigen DNAPolymerasedeltaProcessivityFactor

ProductInformation
Format	Ascites
Presentation	Ascitesfluidcontainingnopreservatives

StorageandShippingInformation
StorageConditions	Maintainfrozenat-20°Cinundilutedaliquotsforupto12monthsfromdateofreceipt.Avoidrepeatedfreeze/thawcycles.

Applications
Application	Anti-PCNAAntibody,clone131-11912isamousemonoclonalantibodyfordetectionofPCNAalsoknownasProliferatingCellNuclearAntigen,DNAPolymerasedeltaProcessivityFactor&hasbeenvalidatedinIF,ICC,IHC(P).
KeyApplications	Immunofluorescence Immunocytochemistry Immunohistochemistry(Paraffin)
ApplicationNotes	Immunohistochemistryonformalinfixed,deparaffinizedtissuesections Frozentissuesections. Smears. Cytospins. DonotuseSeruminblockingsteps. Alsoseehttp://ajp.amjpathol.org/cgi/content/abstract/141/6/1453forimportantreactivityinformationon19A2inparaffinfixedtissues. Finalworkingdilutionsmustbedeterminedbyenduser. IMMUNOHISTOCHEMICALPROCEDUREFORMAB4078 ThefollowinghistochemicalimmunoperoxidaseprotocolhasbeenoptimizedforusewithMAB4078. METHODOLOGY:ThismethodisusedwithstandardstreptavidinorABCimmunoperoxidasemethodsusingaDABsubstraterunatroomtemperaturewithparaffinorfrozensections,smearsorcytospins. NOTE:Optimalfixationtimeforparaffinprocessedtissuefixedin10%NeutralBufferedFormalinis12-14hours. REQUIREDSUPPLEMENTALMATERIAL:* 1.3%HydrogenPeroxide:commerciallyavailable. 2.PBS 3.EthylAlcohol:histologygradeorequivalent 4.Methanol:histologygradeorequivalent 5.Acetone:histologygradeorequivalent 6.Xylene(histologygrade)orXyleneSubstitute(Hemo-D) 7.MountingMedia:Krystalon(Harleco)orequivalent 8.Gill#1Hematoxylin:Sigma#GHS-1-32orequivalent 9.GoatAnti-MouseBiotin 10.StreptavidinHRP(optionalreagent) 11.3,3"Diaminobenzidinetetrahydrochloride(DAB):Sigma#D-5637orequivalent 12.ABC:VectorKit#PK4000orequivalent REAGENTPREPARATION: 1.Primaryantibody:PrepareastocksolutionofantibodyinPBS 2.GoatAnti-MouseBiotin:Approximatedilutionof1:100inPBS(10mg/test) 3.Streptavidin:5mLstocksolutionand995mLPBS(1/200dilution).Letsit15minutesbeforeuse(yieldasavolumesufficientfor~20slides)ORifusingABCkitfromVectorfollowasinstructed. 4.DAB:Reconstitute1.5mgofpowderwith100mLofPBS.Activate15minutespriortousebyadding100mL3%H2O2(yieldsavolumesufficientfor~20slides).WARNING:DABisapossIBLecarcinogen.Avoidcontactwithskin. 5.PBS:Usingagraduatedcylinder,reconstitutewith500mLofdistilledwater. 6.AlcoholBaths:Alldilutionsarealcoholsmadewithdistilledwater. PROCEDURE: PARAFFINSECTIONS (cutat4-5microns,10%NeutralBufferedFormalinfixed): 1.Placeslidesin62°Covenfor15minutes. 2.PlaceslidesinHemo-D,orxylene,for15minutes. 3.De-paraffinizeslidesthroughbathsof:A)xylene/100%ethanol1:1(orHemo-D/100%ethanol1:1),B)100%ethylalcoholx2,C)95%ethylalcohol,D)90%ethylalcohol,E)80%ethylalcohol,F)70%ethylalcoholx2(2minuteseachbath). 4.Placeslidesinrunningcooltapwaterfor5minutes. 5.Shakeoffexcesswater.Placeslidesonincubationracksheldatroomtemperature.CirclesectionswithPAPpen(Newcomer,Inc.)ordiamondpentoisolatesection. 6.Floodsectionswithprimaryantibodyandincubatefor35-40minutes. 7.WipeoffexcessPBS.FloodsectionswithGoatAnti-MouseBiotinreagentandincubatefor35-40minutes. 8.WashwithPBS3-5min 9.WashagainwithPBS3-5min. 10.WipeoffexcessPBS.Floodsectionswithstreptavidin-HRPandincubatefor10minutesORfloodsectionswithABCandincubatefor35-40minutes. 11.WashwithPBS3-5min. 12.WipeoffexcessPBS.FloodsectionswithDABandincubatefor6-8minutes. 13.Placeslidesincoolrunningtapwaterfor5minutes. 14.PlaceslidesinGills#1hematoxylinfor90seconds. 15.Placeslidesincoolrunningtapwaterfor5minutes. 16.DehydrateslidesthroughbathsofA)70%ethylalcohol,B)80%ethylalcohol,C)90%ethylalcohol,D)95%ethylalcoholx2,E)100%ethylalcoholx2,F)xylene/100%ethanol1:1(orHemo-D/100%ethanol1:1)x2,G)xyleneorHemo-Dx2(2minuteseachbath. 17.Mountslidesusingmountingmedia. FROZENSECTIONS(cutat8microns):Pullsectionsfromfreezer(orcutfresh),airdryatroomtemperature(orunderafanfor15minutes),fixedin4oCacetonefor4minutes,airdryfor30minutesatroomtemperature(orunderafanfor15minutes),thenfollowaboveprocedurebeginningat#5. SMEARS:Methanolfixfor30minutesat4oC,thenfollowaboveprocedurebeginningat#5. CYTOSPINS:CelllinesandothercellpopulationspreparedbyresUSPending,ineachwell,~40,000cellsusing500mL1%NormalGoatSerumperwell.Centrifugefor6minutesat1200RPM.Airdryfor30minutesatroomtemperature(orunderafanfor15minutes).Iftumorcellsarebeingscreened,fixinRTacetonefor2-3minutes.Ifperipheralbloodprepsarebeingscreened,fixin100%methanolfor20minutes.Airdrytheslidesfor30minutesatroomtemperature(or15minutesunderafan)andbeginat#5above. NOTE:WhenstainingusingthePCNAantibodiesastheprimaryDONOTuseNormalGoatSerumoranyotherserumblockingstep.

BIOLOGicalInformation
Immunogen	PurifiedPCNA
Clone	131-11912
Host	Mouse
Specificity	SpecificforPCNAa36kDaproliferationassociatedantigen.
Isotype	IgM
SpeciesReactivity	Bovine Canine Human Mouse Rat Rabbit Fish Vertebrates
AntibodyType	MonoclonalAntibody
EntrezGeneNumber	NM_002592.2 NM_182649.1
EntrezGeneSummary	TheproteinencodedbythisgeneisfoundinthenucleusandisacofactorofDNApolymerasedelta.TheencodedproteinactsasahomotrimerandhelpsincreasetheprocessivityofleADIngstrandsynthesisduringDNAreplication.InresponsetoDNAdamage,thisproteinisubiquitinatedandisinvolvedintheRAD6-dependentDNArepairpathway.Twotranscriptvariantsencodingthesameproteinhavebeenfoundforthisgene.Pseudogenesofthisgenehavebeendescribedonchromosome4andontheXchromosome.
GeneSymbol	PCNA Cyclin cyclin MGC8367
UniProtNumber	P12004
UniProtSummary	FUNCTION:SwissProt:P12004#ThisproteinisanauxiliaryproteinofDNApolymerasedeltaandisinvolvedinthecontrolofeukaryoticDNAreplicationbyincreasingthepolymerase"sprocessibilityduringelongationoftheleadingstrand. SIZE:261aminoacids;28769Da SUBUNIT:Homotrimer.InteractswithKCTD10(Bysimilarity).Formsacomplexwithactivator1heteropentamerinthepresenceofATP.InteractswithPOLH,POLK,DNMT1,ERCC5/XPG,FEN1,CDC6,APEX2andPOLDIP2.InteractswithEXO1andSHPRH. SUBCELLULARLOCATION:Nucleus. PTM:Uponmethylmethanesulfonate-inducedDNAdamage,mono-ubiquitinatedbytheUBE2B-RAD18complexonLys-164.Thisinducesnon-canonicalpoly-ubiquitinationonLys-164through"Lys-63"linkageofubiquitinmoietiesbytheE2complexUBE2N-UBE2V2andtheE3ligaseSHPRH,whihisrequiredforDNArepair. DISEASE:SwissProt:P12004#Antibodiesarepresentinserafrompatientswithsystemiclupuserythematosus. SIMILARITY:SwissProt:P12004##BelongstothePCNAfamily.

PhysicochemicalInformation

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Millipore/MAB4078 | Anti-PCNA Antibody, clone 131-11912/MAB4078/100 µL

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