微孔/17-486 |磷酸化IκB（Ser32）STAR ELISA试剂盒/17-486/96分析
市场价：
¥11500.00
美元价：
6900.00
产品分类：
夹心法ELISA
公司分类：
Sandwich_method_ELISA
联系Q Q：
3392242852
电话号码：
4000-520-616
电子邮箱：
info@ebiomall.com
商品介绍
| Description | |
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| CatalogueNumber | 17-486 |
| BrandFamily | Upstate |
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| Description | phospho-IκB(Ser32)STARELISAKit |
| BackgroundInformation | I.TESTPRINCIPLE: ThecolorimetricSTAR(SignalTransductionAssayReaction)ELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtodetectspecificlevelsofsignalingtargetsinwholecellextracts.TheIκBplateiscoatedwithaspecificmousemonoclonalIκBcaptureantibodyonthemicrowellsofthe96-wellclearplate.SamplelysateorthestandardincludedinthekitareincubatedinthemicrowellsallowingIκBantigentobecapturedintheplatewells.Theplateisthenwashedtoremoveanyun-boundnonspecificmaterial.Aspecificrabbitanti-phospho-IκBantibodyisaddedtodetectthecapturedIκBontheplatewellthatisphosphorylatedonSer32.TheunbounddetectionantibodyiswashedawayfollowedbyincubationwithanHRP-conjugatedanti-rabbitantibody.Thisallowsforasensitiveenzymaticdetectionofthesample.AftertheadditionofTMBsubstrateandstopsolutiontheabsorbanceismeasuredat450nmusingaplatereader. Theentireassaytakeslessthan5hourstocompletewithminimalhands-ontime.Manyofthereagentsaresuppliedinready-touseformulationsforeaseofuse.ThekitalsoincludesastandardthatisrunasbothapositivecontrolandtogenerateastandardcurveforIκBmeasurement. II.BACKGROUND: NFκBsignalingisacriticalregulatornotonlyofimmunefunction,butalsoofproliferationversusapoptosisinresponsetovariousstimuli.Inamajorityofunstimulatedcells,theNFκBtranscriptionfactorsexistintheirinactiveformandareretainedinthecytoplasmbytheboundinhibitoryIκBproteinsthatpreventitfromenteringthenucleus,thuskeepingNFκBinaninactivestate.Uponstimulationbymultipleinducersincludingvirusesorcytokines,suchasTNFα,IL-1,orPMA,IκBαisrapidlyphosphorylatedbyIKK(IκBKinase),whichphosphorylatesIκBonserines32and36,allowingrecognitionbytheubiquitinligasemachinery,leADIngtoitspolyubiquinationanddegradation,resultinginthereleaseoftheNFκBcomplex.OnceIκBisdegraded,NFκBisabletoinitiatetranscription.TheNFκBcomplexthentranslocatestothenucleuswhereitactivatesgenetranscription.NFκBinducesthetranscriptionofitsowninhibitor,IκBα,causinganautoregulatorymechanismofNFκBactivityandgeneratingtheinactiveformofNFκB.ThenewlyformednuclearNFκB-IκBαcomplexesarethenexportedouttothecytoplasm,therebyreestablishingthecytoplasmicpoolofinactiveNFκBcomplexesprimedforanotherroundofactivationtotakeplace.ThewidevarietyofgenesregulatedbyNFκBincludesthoseencodingcytokines,chemokines,adhesionmolecules,acutephaseproteins,andinducIBLeeffectorenzymes.MutantIκBinwhichserines32and36arechangedtoalanines,isnotphosphorylated,andthereforenotdegraded.CellsexpressingthisproteinarenotabletoactivateNFκB,providingausefultooltostudytheroleofNFκBinvariouspathwaysandprocesses. |
| MaterialsRequiredbutNotDelivered | 1.Multi-channelorrepeatingPipettes 2.Plateshaker(optional) 3.Pipettors&tipscapableofaccuratelymeasuring1-1000μL 4.GraduatedSEROlogicalpipettes 5.96-wellmicrotiterPlateReaderwith450nmfilter 6.Graphingsoftwareforplottingdataorgraphpaperformanualplottingofdata 7.Microfugetubesforstandardandsampledilutions 8.Mechanicalvortex 9.1litercontainer 10.Distilledordeionizedwater |
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| Detectionmethod | Colorimetric |
| StorageandShippingInformation | |
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| StorageConditions | 1yearat4°Cfromdateofshipment |
| BIOLOGicalInformation | |
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| EntrezGeneNumber | |
| EntrezGeneSummary | NFKB1(MIM164011)orNFKB2(MIM164012)isboundtoREL(MIM164910),RELA(MIM164014),orRELB(MIM604758)toformtheNFKBcomplex.TheNFKBcomplexisinhibitedbyI-kappa-Bproteins(NFKBIAorNFKBIB,MIM604495),whichinactivateNF-kappa-Bbytrappingitinthecytoplasm.PhosphorylationofserineresiduesontheI-kappa-Bproteinsbykinases(IKBKA,MIM600664,orIKBKB,MIM603258)marksthemfordestructionviatheubiquitinationpathway,therebyallowingactivationoftheNF-kappa-Bcomplex.ActivatedNFKBcomplextranslocatesintothenucleusandbindsDNAatkappa-B-bindingmotifssuchas5-primeGGGRNNYYCC3-primeor5-primeHGGARNYYCC3-prime(whereHisA,C,orT;RisanAorGpurine;andYisaCorTpyrimidine).[suppliedbyOMIM] |
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| UniProtSummary | FUNCTION:SwissProt:P25963#InhibitsNF-kappa-Bbycomplexingwithandtrappingitinthecytoplasm.MaybeinvolvedinregulationoftranscriptionalresponsestoNF-kappa-B,includingcelladhesion,immuneandproinflammatoryresponses,apoptosis,differentiationandgrowth.Controlledbysequentialserine-phosphorylation,ubiquitinationanddegradation.Tyrosine-phosphorylationcouldonlyleadtodissociationfromNF-kappa-B. SIZE:317aminoacids;35609Da SUBUNIT:Interactswithp65(RELA).InteractswithNKIRAS1andNKIRAS2.InteractswithHBVproteinX. SUBCELLULARLOCATION:Cytoplasm. PTM:Phosphorylated;disablesinhibitionofNF-kappa-BDNA-bindingactivity.&Ubiquitinated;subsequenttostimulus-dependentphosphorylationonserines,polyubiquitinationtargetstheproteinforrapiddegradationviatheubiquitinsystem. DISEASE:SwissProt:P25963#DefectsinNFKBIAareacauseofautosomaldominantanhidroticectodermaldysplasiawithimmunodeficiency(AD-EDA-ID)[MIM:164008].Ectodermaldysplasias(EDs)constituteaheterogeneousgroupofdevelopmentaldisordersaffectingtissuesofectodermalorigin.EDsarecharacterizedbyabnormaldevelopmentoftwoormoreectodermalstructuressuchashair,teeth,nailsandsweatglands,withorwithoutanyadditionalclinicalsign.Eachcombinationofclinicalfeaturesrepresentsadifferenttypeofectodermaldysplasia. SIMILARITY:SwissProt:P25963##BelongstotheNF-kappa-Binhibitorfamily.&Contains5ANKrepeats. |
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