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Millipore/MAB4072 | Anti-BrdU Antibody, clone 131-14871/MAB4072/100 µg

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貨號: MAB4072

品牌: Millipore

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Description
CatalogueNumber	MAB4072
BrandFamily	Chemicon®
TradeName	Chemicon
Description	Anti-BrdUAntibody,clone131-14871
AlternateNames	BrdU
BackgroundInformation	Thethymidineanalog5-bromo-2′-deoxyuridine(BrdU)isacommonreagentusedforbothcellproliferationassaysandforthedetectionofapoptoticcells.BrdUisreADIlyincorporatedintonewlysynthesizedDNA,labelingcellsthathaveprogressedthroughtheS-phaseofthecellcycleandtherebyservingasaMarkerforproliferatingcells.

ProductInformation
Format	Purified
Control	Brdutreatedcells
Presentation	Liquidin10mMPhosphatebuffer,pH7.4containing150mMNaCland0.1%sodiumazide.

StorageandShippingInformation
StorageConditions	Maintainfor2yearsat-20°Cfromdateofshipment.Aliquottoavoidrepeatedfreezingandthawing.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.

Applications
Application	Anti-BrdUAntibody,clone131-14871isaMouseMonoclonalAntibodyfordetectionofBromodeoxyuridinealsoknownasBrdU&hasbeenvalidatedinFC,IHC,IHC(P).Thisbromodeoxyuridineantibodyisexpecttoreactinallspecies.
KeyApplications	FlowCytometry Immunohistochemistry Immunohistochemistry(Paraffin)
ApplicationNotes	Immunohistochemistryondeparaffinizedsectionsofkidney,duodenum,andlivertissue:1:1000-1:1250. Flowcytometryon100μLof106cells:1:5000. Optimaldilutionsmustbedeterminedbytheenduser. IMMUNOHISTOCHEMICALPROCEDURE FORPARAFFINEMBEDDEDTISSUESECTIONS STAININGPROCEDURE: 1.Deparaffinizesectionsinxylene3x5minutes.ResinembeddedorGMA(Glycolmethacrylate,2-hydroxyethylmethacrylate)treatedsectionsitisunnecessarytodeparaffinizeasthereisnoparaffin. 2.Placeslidesingradedalcohols(100,95,90%)towaterat2minuteintervals,andairdry.(OmitforGMAsections) 3.CirclesectionswithPAPpenordiamondpentoidentifyanddrythoroughly. 4.ForGMAorresinsections,placeslidesin0.5%Tween/PBS(pH7.6)for29minutes. 5.Incubateinprewarmed(40°C)1NHClfor1hour. 6.Wash3x3minuteswithPBS(pH7.6).Slidesmaybeheldatthispointandtheprocedurecontinuedthefollowingday. 7.Incubatein1XTrypsinsolution(0.02-0.05%w/v,prewarmedto40°C)for20minutesatroomtemperatureforNBF(normalbufferedformalin)fixedtissueor10minutesforCarnoy"sfixedtissue. 8.Wash3x5minuteswithPBS(pH7.6). 9.Incubatesectionsin1%H2O2(10mL30%H2O2in290mLmethanol)for20minutes,orGMAsectionsin3%H2O2(10mL30%H2O2in90mLwater)for3minutes. 10.Wash2x2minuteswithPBS(pH7.6). 11.UsingtheVectorABCkit,add3dropsofnormalhorseserumto10mLofPBS.Usethissolutiontoblocktheslidesfor20minutesatroomtemperature. 12.BlottheslidesofexcessnormalhorseserumandincubatewithMAB4072dilutedinPBS/BSA/Tween20for1houratroomtemperature. 13.Wash3x3minuteswithPBS(pH7.6). 14.Add3dropsofnormalhorseserumto10mLofPBSandthenadd1dropofbiotinylatedantibodystock.Incubatetheslideswiththisdilutedsecondantibodyfor30minutesatroomtemperature. 15.PreparetheVectastainABCReagentbyadding2dropsofreagentAto5mLofPBS.Thenadd2dropsofReagentBtothesamemixingbottle.Allowtositforabout30minutespriortouse. 16.Wash3x3minuteswithPBS(pH7.6). 17.IncubatetheslideswithABCReagentfor30minutesatroomtemperature. 18.Wash3x3minuteswithPBS(pH7.6). 19.Preparethesubstratebymixinganequalvolumeof0.02%H2O2(madeindistilledwaterfroma30%stocksolution)and0.1%DABmadein0.1MTrisbuffer,pH7.2.TheH2O2shouldbefreshlypreparedfromconcentratedstock.BecausemanyperoxidesubstratesareunstableinthepresenceofH2O2orwhenexposedtolight,thesubstrateshouldbepreparedjustpriortouse.SinceDABisasUSPectedcarcinogen,careshouldbetakeninhandlinganddisposingofallperoxidasesubstrates. 20.StaintheslideswithDABsubstratefor2-5minutes. 21.Wash2x2minuteswithdistilledwater. 22.Counterstainwithhemotoxylin,dehydratethroughxylene,andmountthecoverslip.

BIOLOGicalInformation
Clone	131-14871
Concentration	PleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
Host	Mouse
Specificity	RecognizesBrdU(Bromodeoxyuridine).
Isotype	IgG1
SpeciesReactivity	All
AntibodyType	MonoclonalAntibody
PurificationMethod	ProteinAPurfied

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Millipore/MAB4072 | Anti-BrdU Antibody, clone 131-14871/MAB4072/100 µg

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