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Millipore/17-484 | phospho-IR (β Subunit) (Tyr 1162/1163) STAR ELISA Assay Kit/17-484/96 assays

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貨號: 17-484

品牌: Millipore

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Description
CatalogueNumber	17-484
BrandFamily	Upstate
TradeName	STAR Upstate
Description	phospho-IR(βSubunit)(Tyr1162/1163)STARELISAAssayKit
AlternateNames	InsulinReceptor IR
BackgroundInformation	I.TESTPRINCIPLE TheUPSTATE?colorimetricSTAR(SignalTransductionAssayReaction)ELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtodetectspecificlevelsofsignalingtargetsinwholecellextracts.TheIRplateiscoatedwithaspecificmousemonoclonalIR(βsubunit)captureantibodyonthemicrowellsofthe96-wellclearplate.SamplelysateorthestandardsincludedinthekitareincubatedinthemicrowellsallowingIRtobecapturedintheplatewells.Theplateisthenwashedtoremoveanyunboundnon-specificmaterial.Thewellsarethenincubatedwithaspecificrabbitanti-phospho-IRantibodytodetectthecapturedIRphosphorylatedonTyrosines1162and1163.TheunbounddetectionantibodyiswashedawayfollowedbyincubationwithanHRP-conjugatedanti-rabbitantibody.Thisallowsforasensitiveenzymaticdetectionofthesample.AftertheadditionofTMBsubstrateandstopsolutiontheabsorbanceismeasuredat450nmusingaplatereader. Theentireassaytakeslessthan5hourstocompletewithminimalhands-ontime.Manyofthereagentsaresuppliedinready-touseformulationsforeaseofuse.Thekitalsoincludesastandardthatisrunasbothapositivecontrolandtodevelopastandardcurve. II.IRBACKGROUND TheInsulinReceptor(IR)issynthesizedasasinglepolypeptide,whichissubsequentlycleavedtogenerateanextracellularαchainandatransmembraneandintracellularβchain,tetheredtogetherbydisulfidebonds.Theβchainhasmultipletyrosinephosphorylationsites,includingthreeautophosphorylationsitesatitsactivationloop.Insulinstimulatesthesignalingcascadebybindingtotheinsulinreceptorthroughitsextracellularαsubunits.Thisstimulatesthetyrosinekinaseactivityoftheβsubunitsofthereceptor.ThereceptorthenautophosphorylatesitselfandphosphorylatestheIRS(InsulinReceptorSubstrate)proteinsthatareveryimportantmodulatorsofinsulinsignaling.PhosphorylatedIRS,inturn,interactswithseveraldownstreameffectorsofinsulinsignaling,includingPI3Kinasep85,GRB2,SHP2,Nck,Crk,andFyn.TheoverallstructureoftheIRishighlyhomologoustotheIGF-IReceptor,exceptintheirc-termini,wherethetwoproteinsdivergesomewhat.InsulinsignalingishighlydependentonthePI3KinasepathwayandAkt,whichappeartomediatethefunctionsofinsulin.
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

ProductInformation
Components	CapturePlatepre-coatedwithanti-IRantibody:(PartNo.17-483A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch. Anti-phospho-IR(Tyr1162/1163)detectionantibody:(PartNo.17-483B)Onebottle(11mL)ofanti-phospho-IR(Tyr1162/1163)detectionantibodycontainingsodiumazide,readytouse. ELISADiluent:(PartNo.17-483C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse. 25XELISAWashBuffer:(PartNo.17-483D)Onebottle(50mL)of25XELISAWashBuffer. Anti-RabbitIgGHRPconjugate:(PartNo.17-483E)Onevial(125μL)of100Xanti-rabbitHRPconjugatecontainingthimerosal. HRPDiluent:(PartNo.17-483F)Onebottle(25mL)ofHRPDiluentcontainingthimerosal. TMBSolution:(PartNo.17-483G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse. StopSolution:(PartNo.17-483H)Onebottle(25mL)ofstopsolution,readytouse. phospho-IRStandard:(PartNo.17-483I)TwovialsofIR((Tyr1162/1163)standard,lyophilized. PlateCovers:Twoplatecovers.
Detectionmethod	Chromogenic

ProductInformation

Components

CapturePlatepre-coatedwithanti-IRantibody:(PartNo.17-483A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch.
Anti-phospho-IR(Tyr1162/1163)detectionantibody:(PartNo.17-483B)Onebottle(11mL)ofanti-phospho-IR(Tyr1162/1163)detectionantibodycontainingsodiumazide,readytouse.
ELISADiluent:(PartNo.17-483C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse.
25XELISAWashBuffer:(PartNo.17-483D)Onebottle(50mL)of25XELISAWashBuffer.
Anti-RabbitIgGHRPconjugate:(PartNo.17-483E)Onevial(125μL)of100Xanti-rabbitHRPconjugatecontainingthimerosal.
HRPDiluent:(PartNo.17-483F)Onebottle(25mL)ofHRPDiluentcontainingthimerosal.
TMBSolution:(PartNo.17-483G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse.
StopSolution:(PartNo.17-483H)Onebottle(25mL)ofstopsolution,readytouse.
phospho-IRStandard:(PartNo.17-483I)TwovialsofIR((Tyr1162/1163)standard,lyophilized.
PlateCovers:Twoplatecovers.

Detectionmethod

Chromogenic

StorageandShippingInformation
StorageConditions	Maintaintheunopenedkitat2-8°Cuntilexpirationdate.

Applications
Application	Thisphospho-IR(βSubunit)(Tyr1162/1163)STARELISAAssayKitisusedtomeasure&quantifyInsulinlevels.
KeyApplications	ELISA

BIOLOGicalInformation
SpeciesReactivity	Human Mouse Rat
AnalytesAvailable	Insulin
EntrezGeneNumber	NM_000208.2 NM_001079817.1
EntrezGeneSummary	Afterremovaloftheprecursorsignalpeptide,theinsulinreceptorprecursorispost-translationallycleavedintotwochains(alphaandbeta)thatarecovalentlylinked.Bindingofinsulintotheinsulinreceptor(INSR)stimulatesglucoseuptake.Twotranscriptvariantsencodingdifferentisoformshavebeenfoundforthisgene.
GeneSymbol	INSR CD220 IR HHF5
Modifications	Phosphorylation
UniProtNumber	P06213
UniProtSummary	FUNCTION:SwissProt:P06213#Thisreceptorbindsinsulinandhasatyrosine-proteinkinaseactivity.IsoformShorthasahigheraffinityforinsulin.Mediatesthemetabolicfunctionsofinsulin.BindingtoinsulinstimulatesassociationofthereceptorwithdownstreammediatorsincludingIRS1andphosphatidylinositol3"-kinase(PI3K).CanactivatePI3Keitherdirectlybybindingtothep85regulatorysubunit,orindirectlyviaIRS1. SIZE:1382aminoacids;156307Da SUBUNIT:Tetramerof2alphaand2betachainslinkedbydisulfidebonds.Thealphachainscontributetotheformationoftheligand-bindingdomain,whilethebetachainscarrythekinasedomain.InteractswithSORBS1butdissociatesfromitfollowinginsulinstimulation.BindsSH2B2.InteractswiththePTB/PIDdomainsofIRS1andSHC1invitrowhenautophosphorylatedontyrosineresidues.ThesequencessurroundingthephosphorylatedNPXYmotifcontributedifferentiallytoeitherIRS1orSHC1recognition.InteractswiththeSH2domainsofthe85kDaregulatorysubunitofPI3K(PIK3R1)invitro,whenautophosphorylatedontyrosineresidues.InteractswithSOCS7. SUBCELLULARLOCATION:Membrane;Single-passtypeImembraneprotein. TISSUESPECIFICITY:IsoformLongandisoformShortareexpressedintheperipheralnerve,kidney,liver,striatedmuscle,fibroblastsandskin.IsoformShortisexpressedalsointhespleenandlymphoblasts. PTM:AfterbeingtransportedfromtheendoplasmicreticulumtotheGolgiapparatus,thesingleglycosylatedprecursorisfurtherglycosylatedandthencleaved,followedbyitstransporttotheplasmamembrane.&Autophosphorylatedontyrosineresiduesinresponsetoinsulin.&PhosphorylationofTyr-999isrequiredforIRS1-andSHC1-binding. DISEASE:SwissProt:P06213#DefectsinINSRarethecauseofinsulinresistance(Insresistance)[MIM:125853].&DefectsinINSRarethecauseofRabson-Mendenhallsyndrome[MIM:262190];alsoknownasMendenhallsyndrome.Itisasevereinsulinresistancesyndromecharacterizedbyinsulin-resistantdiabetesmellituswithpinealhyperplasiaandsomaticabnormalities.Typicalfeaturesincludecoarse,senile-appearingfacies,dentalandskinabnormalities,aBDominaldistension,andphallicenlargement.Inheritanceisautosomalrecessive.&DefectsinINSRarethecauseofleprechaunism[MIM:246200];alsoknownasDonohuesyndrome.Leprechaunismrepresentsthemostsevereformofinsulinresistancesyndrome,characterizedbyintrauterineandpostnatalgrowthretardationanddeathinearlyinfancy.Inheritanceisautosomalrecessive.&DefectsinINSRmaybeassociatedwithnoninsulin-dependentdiabetesmellitus(NIDDM)[MIM:125853];alsoknownasdiabetesmellitustype2.&DefectsinINSRarethecauseoffamilialhyperinsulinemichypoglycemia5(HHF5)[MIM:609968].Familialhyperinsulinemichypoglycemia[MIM:256450],alsoreferredtoascongenitalhyperinsulinism,nesidioblastosis,orpersistenthyperinsulinemichypoglycemiaofinfancy(PPHI),isthemostcommoncauseofpersistenthypoglycemiaininfancyandisduetodefectivenegativefeedbackregulationofinsulinsecretionbylowglucoselevels.&DefectsinINSRarethecauseofinsulin-resistantdiabetesmellituswithacanthosisnigricanstypeA(IRANtypeA)[MIM:610549].Thissyndromeischaracterizedbytheassociationofsevereinsulinresistance(manifestedbymarkedhyperinsulinemiaandafailuretorespondtoexogenousinsulin)withtheskinlesionacanthosisnigricansandovarianhyperandrogenisminadolescentfemalesubjects.Womenfrequentlypresentwithhirsutism,acne,amenorrheaoroligomenorrhea,andvirilization.ThissyndromeisdifferentfromthetypeBthathasbeendemonstratedtobesecondarytothepresenceofcirculatingautoantibodiesagainsttheinsulinreceptor. SIMILARITY:SwissProt:P06213##Belongstotheproteinkinasesuperfamily.Tyrproteinkinasefamily.Insulinreceptorsubfamily.&Contains2fibronectintype-IIIdomains.&Contains1proteinkinasedomain.

PhysicochemicalInformation
Sensitivity	Sensitivity:1Unit/mL RangeofDetection:1.6-100Units/mL

Dimensions

MaterialsInformation

MaterialsInformation

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