biologicalkb936929
kb936929 时间:2021-02-28 阅读:()
ProductnameAnti-NF-kBp65antibodyDescriptionChickenpolyclonaltoNF-kBp65HostspeciesChickenTestedapplicationsSuitablefor:WBSpeciesreactivityReactswith:Rat,HumanPredictedtoworkwith:Mouse,Cow,Pig,Chimpanzee,Macaquemonkey,Gorilla,ChinesehamsterImmunogenSyntheticpeptidewithinHumanNF-kBp65aa500totheC-terminusconjugatedtokeyholelimpethaemocyanin.
Theexactsequenceisproprietary.
PositivecontrolThisantibodygaveapositivesignalinthefollowinglysates:HeLawholecell;Jurkatwholecell;Ratspleentissue.
GeneralnotesReproducibilityiskeytoadvancingscientificdiscoveryandacceleratingscientists'nextbreakthrough.
Abcamisleadingthewaywithourrangeofrecombinantantibodies,knockout-validatedantibodiesandknockoutcelllines,allofwhichsupportimprovedreproducibility.
Wearealsoplanningtoinnovatethewayinwhichwepresentrecommendedapplicationsandspeciesonourproductdatasheets,sothatonlyapplications&speciesthathavebeentestedinourownlabs,oursuppliersorbyselectedtrustedcollaboratorsarecoveredbyourAbpromiseguarantee.
Inpreparationforthis,wehavestartedtoupdatetheapplications&speciesthatthisproductisAbpromiseguaranteedfor.
Wearealsoupdatingtheapplications&speciesthatthisproducthasbeen"predictedtoworkwith,"howeverthisinformationisnotcoveredbyourAbpromiseguarantee.
Applications&speciesfrompublicationsandAbreviewsthathavenotbeentestedinourownlabsorinthoseofoursuppliersarenotcoveredbytheAbpromiseguarantee.
Pleasecheckthatthisproductmeetsyourneedsbeforepurchasing.
Ifyouhaveanyquestions,specialrequirementsorconcerns,pleasesendusaninquiryand/orcontactourSupportteamaheadofpurchase.
Recommendedalternativesforthisproductcanbefoundbelow,aswellascustomerreviewsandQ&As.
ProductdatasheetProductdatasheetAnti-NF-kBp65antibodyab140751Anti-NF-kBp65antibodyab1407512Abreviews2References2ImagesOverview1FormLiquidStorageinstructionsShippedat4°C.
Upondeliveryaliquot.
Storeat-20°Cor-80°C.
Avoidfreeze/thawcycle.
StoragebufferpH:7.
40Preservative:0.
02%SodiumazideConstituents:PBS,3%BSAPurityProteinLpurifiedClonalityPolyclonalIsotypeIgYFunctionNF-kappa-Bisapleiotropictranscriptionfactorwhichispresentinalmostallcelltypesandisinvolvedinmanybiologicalprocessedsuchasinflammation,immunity,differentiation,cellgrowth,tumorigenesisandapoptosis.
NF-kappa-Bisahomo-orheterodimericcomplexformedbytheRel-likedomain-containingproteinsRELA/p65,RELB,NFKB1/p105,NFKB1/p50,RELandNFKB2/p52andtheheterodimericp65-p50complexappearstobemostabundantone.
Thedimersbindatkappa-BsitesintheDNAoftheirtargetgenesandtheindividualdimershavedistinctpreferencesfordifferentkappa-Bsitesthattheycanbindwithdistinguishableaffinityandspecificity.
Differentdimercombinationsactastranscriptionalactivatorsorrepressors,respectively.
NF-kappa-Biscontrolledbyvariousmechanismsofpost-translationalmodificationandsubcellularcompartmentalizationaswellasbyinteractionswithothercofactorsorcorepressors.
NF-kappa-BcomplexesareheldinthecytoplasminaninactivestatecomplexedwithmembersoftheNF-kappa-Binhibitor(I-kappa-B)family.
Inaconventionalactivationpathway,I-kappa-BisphosphorylatedbyI-kappa-Bkinases(IKKs)inresponsetodifferentactivators,subsequentlydegradedthusliberatingtheactiveNF-kappa-Bcomplexwhichtranslocatestothenucleus.
NF-kappa-Bheterodimericp65-p50andp65-c-Relcomplexesaretranscriptionalactivators.
TheNF-kappa-Bp65-p65complexappearstobeinvolvedininvasin-mediatedactivationofIL-8expression.
TheinhibitoryeffectofI-kappa-BuponNF-kappa-Bthecytoplasmisexertedprimarilythroughtheinteractionwithp65.
p65showsaweakDNA-bindingsitewhichcouldcontributedirectlytoDNAbindingintheNF-kappa-Bcomplex.
AssociateswithchromatinattheNF-kappa-BpromoterregionviaassociationwithDDX1.
SequencesimilaritiesContains1RHD(Rel-like)domain.
Domainthe9aaTADmotifisatransactivationdomainpresentinalargenumberofyeastandanimaltranscriptionfactors.
PropertiesApplicationsOurAbpromiseguaranteecoverstheuseofab140751inthefollowingtestedapplications.
Theapplicationnotesincluderecommendedstartingdilutions;optimaldilutions/concentrationsshouldbedeterminedbytheenduser.
ApplicationAbreviewsNotesWBUseaconcentrationof1g/ml.
Detectsabandofapproximately65kDa(predictedmolecularweight:65kDa).
Target2Post-translationalmodificationsUbiquitinated,leadingtoitsproteasomaldegradation.
DegradationisrequiredforterminationofNF-kappa-Bresponse.
MonomethylatedatLys-310bySETD6.
MonomethylationatLys-310isrecognizedbytheANKrepeatsofEHMT1andpromotestheformationofrepressedchromatinattargetgenes,leadingtodown-regulationofNF-kappa-Btranscriptionfactoractivity.
PhosphorylationatSer-311disruptstheinteractionwithEHMT1withoutpreventingmonomethylationatLys-310andrelievestherepressionoftargetgenes.
PhosphorylationatSer-311disruptstheinteractionwithEHMT1andpromotestranscriptionfactoractivity(Bysimilarity).
PhosphorylationonSer-536stimulatesacetylationonLys-310andinteractionwithCBP;thephosphorylatedandacetylatedformsshowenhancedtranscriptionalactivity.
Reversiblyacetylated;theacetylationseemstobemediatedbyCBP,thedeacetylationbyHDAC3.
AcetylationatLys-122enhancesDNAbindingandimpairsassociationwithNFKBIA.
AcetylationatLys-310isrequiredforfulltranscriptionalactivityintheabsenceofeffectsonDNAbindingandNFKBIAassociation.
AcetylationcanalsolowerDNA-bindingandresultsinnuclearexport.
InteractionwithBRMS1promotesdeacetylationof'Lys-310'.
CellularlocalizationNucleus.
Cytoplasm.
Nuclear,butalsofoundinthecytoplasminaninactiveformcomplexedtoaninhibitor(I-kappa-B).
ColocalizedwithRELAinthenucleusuponTNF-alphainduction.
Westernblot-Anti-NF-kBp65antibody(ab140751)Lane1:Wild-typeHAP1celllysate(20g)Lane2:NF-κBp60knockoutHAP1celllysate(20g)Lane3:HeLacelllysate(20g)Lane4:A431celllysate(20g)Lanes1-4:Mergedsignal(redandgreen).
Green-ab140751observedat70kDa.
Red-ab8245loadingcontrol,observedat37kDa.
ab140751wasshowntospecificallyreactwithNF-κBp65inwild-typeHAP1cells.
NobandwasobservedwhenNF-κBp65knockoutsampleswereused.
Wild-typeandNF-κBp65knockoutsamplesweresubjectedtoSDS-PAGE.
ab140751(NF-κBp65)andab8245(loadingcontroltoGAPDH)werediluted1μg/mLand1/2000andincubatedovernightat4°C.
BlotsweredevelopedwithIRDye800CWDonkeyanti-ChickenIgG(H+L)andDonkeyAnti-RabbitIgGH&L(IRDye680RD)preadsorbedab216779secondaryantibodiesat1/10000dilutionfor1hatroomtemperaturebeforeimaging.
Images3Westernblot-Anti-NF-kBp65antibody(ab140751)Alllanes:Anti-NF-kBp65antibody(ab140751)at1g/mlLane1:HeLa(Humanepithelialcarcinomacellline)WholeCellLysateLane2:Jurkat(HumanTcelllymphoblast-likecellline)WholeCellLysateLane3:Spleen(Rat)TissueLysateLysates/proteinsat20gperlane.
SecondaryAlllanes:GoatAnti-ChickenIgYH&L(HRP)(ab6877)at1/10000dilutionDevelopedusingtheECLtechnique.
Performedunderreducingconditions.
Predictedbandsize:65kDaObservedbandsize:65kDaAdditionalbandsat:120kDa.
Weareunsureastotheidentityoftheseextrabands.
Exposuretime:1minutePleasenote:Allproductsare"FORRESEARCHUSEONLY.
NOTFORUSEINDIAGNOSTICPROCEDURES"OurAbpromisetoyou:QualityguaranteedandexperttechnicalsupportReplacementorrefundforproductsnotperformingasstatedonthedatasheetValidfor12monthsfromdateofdeliveryResponsetoyourinquirywithin24hoursWeprovidesupportinChinese,English,French,German,JapaneseandSpanishExtensivemulti-mediatechnicalresourcestohelpyouWeinvestigateallqualityconcernstoensureourproductsperformtothehigheststandardsIftheproductdoesnotperformasdescribedonthisdatasheet,wewillofferarefundorreplacement.
ForfulldetailsoftheAbpromise,pleasevisithttps://www.
abcam.
com/abpromiseorcontactourtechnicalteam.
4TermsandconditionsGuaranteeonlyvalidforproductsboughtdirectfromAbcamoroneofourauthorizeddistributors5
Theexactsequenceisproprietary.
PositivecontrolThisantibodygaveapositivesignalinthefollowinglysates:HeLawholecell;Jurkatwholecell;Ratspleentissue.
GeneralnotesReproducibilityiskeytoadvancingscientificdiscoveryandacceleratingscientists'nextbreakthrough.
Abcamisleadingthewaywithourrangeofrecombinantantibodies,knockout-validatedantibodiesandknockoutcelllines,allofwhichsupportimprovedreproducibility.
Wearealsoplanningtoinnovatethewayinwhichwepresentrecommendedapplicationsandspeciesonourproductdatasheets,sothatonlyapplications&speciesthathavebeentestedinourownlabs,oursuppliersorbyselectedtrustedcollaboratorsarecoveredbyourAbpromiseguarantee.
Inpreparationforthis,wehavestartedtoupdatetheapplications&speciesthatthisproductisAbpromiseguaranteedfor.
Wearealsoupdatingtheapplications&speciesthatthisproducthasbeen"predictedtoworkwith,"howeverthisinformationisnotcoveredbyourAbpromiseguarantee.
Applications&speciesfrompublicationsandAbreviewsthathavenotbeentestedinourownlabsorinthoseofoursuppliersarenotcoveredbytheAbpromiseguarantee.
Pleasecheckthatthisproductmeetsyourneedsbeforepurchasing.
Ifyouhaveanyquestions,specialrequirementsorconcerns,pleasesendusaninquiryand/orcontactourSupportteamaheadofpurchase.
Recommendedalternativesforthisproductcanbefoundbelow,aswellascustomerreviewsandQ&As.
ProductdatasheetProductdatasheetAnti-NF-kBp65antibodyab140751Anti-NF-kBp65antibodyab1407512Abreviews2References2ImagesOverview1FormLiquidStorageinstructionsShippedat4°C.
Upondeliveryaliquot.
Storeat-20°Cor-80°C.
Avoidfreeze/thawcycle.
StoragebufferpH:7.
40Preservative:0.
02%SodiumazideConstituents:PBS,3%BSAPurityProteinLpurifiedClonalityPolyclonalIsotypeIgYFunctionNF-kappa-Bisapleiotropictranscriptionfactorwhichispresentinalmostallcelltypesandisinvolvedinmanybiologicalprocessedsuchasinflammation,immunity,differentiation,cellgrowth,tumorigenesisandapoptosis.
NF-kappa-Bisahomo-orheterodimericcomplexformedbytheRel-likedomain-containingproteinsRELA/p65,RELB,NFKB1/p105,NFKB1/p50,RELandNFKB2/p52andtheheterodimericp65-p50complexappearstobemostabundantone.
Thedimersbindatkappa-BsitesintheDNAoftheirtargetgenesandtheindividualdimershavedistinctpreferencesfordifferentkappa-Bsitesthattheycanbindwithdistinguishableaffinityandspecificity.
Differentdimercombinationsactastranscriptionalactivatorsorrepressors,respectively.
NF-kappa-Biscontrolledbyvariousmechanismsofpost-translationalmodificationandsubcellularcompartmentalizationaswellasbyinteractionswithothercofactorsorcorepressors.
NF-kappa-BcomplexesareheldinthecytoplasminaninactivestatecomplexedwithmembersoftheNF-kappa-Binhibitor(I-kappa-B)family.
Inaconventionalactivationpathway,I-kappa-BisphosphorylatedbyI-kappa-Bkinases(IKKs)inresponsetodifferentactivators,subsequentlydegradedthusliberatingtheactiveNF-kappa-Bcomplexwhichtranslocatestothenucleus.
NF-kappa-Bheterodimericp65-p50andp65-c-Relcomplexesaretranscriptionalactivators.
TheNF-kappa-Bp65-p65complexappearstobeinvolvedininvasin-mediatedactivationofIL-8expression.
TheinhibitoryeffectofI-kappa-BuponNF-kappa-Bthecytoplasmisexertedprimarilythroughtheinteractionwithp65.
p65showsaweakDNA-bindingsitewhichcouldcontributedirectlytoDNAbindingintheNF-kappa-Bcomplex.
AssociateswithchromatinattheNF-kappa-BpromoterregionviaassociationwithDDX1.
SequencesimilaritiesContains1RHD(Rel-like)domain.
Domainthe9aaTADmotifisatransactivationdomainpresentinalargenumberofyeastandanimaltranscriptionfactors.
PropertiesApplicationsOurAbpromiseguaranteecoverstheuseofab140751inthefollowingtestedapplications.
Theapplicationnotesincluderecommendedstartingdilutions;optimaldilutions/concentrationsshouldbedeterminedbytheenduser.
ApplicationAbreviewsNotesWBUseaconcentrationof1g/ml.
Detectsabandofapproximately65kDa(predictedmolecularweight:65kDa).
Target2Post-translationalmodificationsUbiquitinated,leadingtoitsproteasomaldegradation.
DegradationisrequiredforterminationofNF-kappa-Bresponse.
MonomethylatedatLys-310bySETD6.
MonomethylationatLys-310isrecognizedbytheANKrepeatsofEHMT1andpromotestheformationofrepressedchromatinattargetgenes,leadingtodown-regulationofNF-kappa-Btranscriptionfactoractivity.
PhosphorylationatSer-311disruptstheinteractionwithEHMT1withoutpreventingmonomethylationatLys-310andrelievestherepressionoftargetgenes.
PhosphorylationatSer-311disruptstheinteractionwithEHMT1andpromotestranscriptionfactoractivity(Bysimilarity).
PhosphorylationonSer-536stimulatesacetylationonLys-310andinteractionwithCBP;thephosphorylatedandacetylatedformsshowenhancedtranscriptionalactivity.
Reversiblyacetylated;theacetylationseemstobemediatedbyCBP,thedeacetylationbyHDAC3.
AcetylationatLys-122enhancesDNAbindingandimpairsassociationwithNFKBIA.
AcetylationatLys-310isrequiredforfulltranscriptionalactivityintheabsenceofeffectsonDNAbindingandNFKBIAassociation.
AcetylationcanalsolowerDNA-bindingandresultsinnuclearexport.
InteractionwithBRMS1promotesdeacetylationof'Lys-310'.
CellularlocalizationNucleus.
Cytoplasm.
Nuclear,butalsofoundinthecytoplasminaninactiveformcomplexedtoaninhibitor(I-kappa-B).
ColocalizedwithRELAinthenucleusuponTNF-alphainduction.
Westernblot-Anti-NF-kBp65antibody(ab140751)Lane1:Wild-typeHAP1celllysate(20g)Lane2:NF-κBp60knockoutHAP1celllysate(20g)Lane3:HeLacelllysate(20g)Lane4:A431celllysate(20g)Lanes1-4:Mergedsignal(redandgreen).
Green-ab140751observedat70kDa.
Red-ab8245loadingcontrol,observedat37kDa.
ab140751wasshowntospecificallyreactwithNF-κBp65inwild-typeHAP1cells.
NobandwasobservedwhenNF-κBp65knockoutsampleswereused.
Wild-typeandNF-κBp65knockoutsamplesweresubjectedtoSDS-PAGE.
ab140751(NF-κBp65)andab8245(loadingcontroltoGAPDH)werediluted1μg/mLand1/2000andincubatedovernightat4°C.
BlotsweredevelopedwithIRDye800CWDonkeyanti-ChickenIgG(H+L)andDonkeyAnti-RabbitIgGH&L(IRDye680RD)preadsorbedab216779secondaryantibodiesat1/10000dilutionfor1hatroomtemperaturebeforeimaging.
Images3Westernblot-Anti-NF-kBp65antibody(ab140751)Alllanes:Anti-NF-kBp65antibody(ab140751)at1g/mlLane1:HeLa(Humanepithelialcarcinomacellline)WholeCellLysateLane2:Jurkat(HumanTcelllymphoblast-likecellline)WholeCellLysateLane3:Spleen(Rat)TissueLysateLysates/proteinsat20gperlane.
SecondaryAlllanes:GoatAnti-ChickenIgYH&L(HRP)(ab6877)at1/10000dilutionDevelopedusingtheECLtechnique.
Performedunderreducingconditions.
Predictedbandsize:65kDaObservedbandsize:65kDaAdditionalbandsat:120kDa.
Weareunsureastotheidentityoftheseextrabands.
Exposuretime:1minutePleasenote:Allproductsare"FORRESEARCHUSEONLY.
NOTFORUSEINDIAGNOSTICPROCEDURES"OurAbpromisetoyou:QualityguaranteedandexperttechnicalsupportReplacementorrefundforproductsnotperformingasstatedonthedatasheetValidfor12monthsfromdateofdeliveryResponsetoyourinquirywithin24hoursWeprovidesupportinChinese,English,French,German,JapaneseandSpanishExtensivemulti-mediatechnicalresourcestohelpyouWeinvestigateallqualityconcernstoensureourproductsperformtothehigheststandardsIftheproductdoesnotperformasdescribedonthisdatasheet,wewillofferarefundorreplacement.
ForfulldetailsoftheAbpromise,pleasevisithttps://www.
abcam.
com/abpromiseorcontactourtechnicalteam.
4TermsandconditionsGuaranteeonlyvalidforproductsboughtdirectfromAbcamoroneofourauthorizeddistributors5
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