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主营：分子类，蛋白类，抗体类，生化类试剂
℡ 4000-520-616
℡ 4000-520-616
Millipore/MAB4078 | Anti-PCNA Antibody, clone 131-11912/MAB4078/100 µL
产品编号：MAB4078
市  场 价：¥5960.00
场      地：美国(厂家直采)
产品分类： 抗体类>一抗>功能性抗体>
联系QQ：1570468124
电话号码：4000-520-616
邮      箱： info@ebiomall.com
美  元  价：$298.00
品      牌： Millipore
公      司：Millipore
公司分类：
Millipore/MAB4078 | Anti-PCNA Antibody, clone 131-11912/MAB4078/100 µL
商品介绍
Description
CatalogueNumberMAB4078
BrandFamilyChemicon®
TradeName
  • Chemicon
DescriptionAnti-PCNAAntibody,clone131-11912
AlternateNames
  • ProliferatingCellNuclearAntigen
  • DNAPolymerasedeltaProcessivityFactor
ProductInformation
FormatAscites
PresentationAscitesfluidcontainingnopreservatives
StorageandShippingInformation
StorageConditionsMaintainfrozenat-20°Cinundilutedaliquotsforupto12monthsfromdateofreceipt.Avoidrepeatedfreeze/thawcycles.
Applications
ApplicationAnti-PCNAAntibody,clone131-11912isamousemonoclonalantibodyfordetectionofPCNAalsoknownasProliferatingCellNuclearAntigen,DNAPolymerasedeltaProcessivityFactor&hasbeenvalidatedinIF,ICC,IHC(P).
KeyApplications
  • Immunofluorescence
  • Immunocytochemistry
  • Immunohistochemistry(Paraffin)
ApplicationNotesImmunohistochemistryonformalinfixed,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
ImmunogenPurifiedPCNA
Clone131-11912
HostMouse
SpecificitySpecificforPCNAa36kDaproliferationassociatedantigen.
IsotypeIgM
SpeciesReactivity
  • Bovine
  • Canine
  • Human
  • Mouse
  • Rat
  • Rabbit
  • Fish
  • Vertebrates
AntibodyTypeMonoclonalAntibody
EntrezGeneNumber
EntrezGeneSummaryTheproteinencodedbythisgeneisfoundinthenucleusandisacofactorofDNApolymerasedelta.TheencodedproteinactsasahomotrimerandhelpsincreasetheprocessivityofleADIngstrandsynthesisduringDNAreplication.InresponsetoDNAdamage,thisproteinisubiquitinatedandisinvolvedintheRAD6-dependentDNArepairpathway.Twotranscriptvariantsencodingthesameproteinhavebeenfoundforthisgene.Pseudogenesofthisgenehavebeendescribedonchromosome4andontheXchromosome.
GeneSymbol
  • PCNA
  • Cyclin
  • cyclin
  • MGC8367
UniProtNumber
UniProtSummaryFUNCTION: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
Dimensions
MaterialsInformation
MaterialsInformation
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