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Millipore/17-462 | Phospho-EGFR (Tyr1068) STAR Assay ELISA Kit/17-462/96 assays

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貨號: 17-462

品牌: Millipore

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Description
CatalogueNumber	17-462
BrandFamily	Upstate
TradeName	STAR Upstate
Description	Phospho-EGFR(Tyr1068)STARAssayELISAKit
AlternateNames	Epidermalgrowthfactorreceptor Proto-oncogenec-ErbB-1 Receptortyrosine-proteinkinaseerbB-1 EGFReceptor EGFR ERBB ERBB2
BackgroundInformation	I.TESTPRINCIPLE TheUPSTATE?colorimetricSTAR(SignalTransductionAssayReaction)ELISAkitisasolidphasesandwichenzymelinkedimmunosorbentassaythatprovidesafast,sensitivemethodtodetectspecificlevelsofsignalingtargetsinwholecellextracts.TheEGFRplateiscoatedwithaspecificmousemonoclonalEGFRcaptureantibody.SamplelysateorthestandardincludedinthekitareincubatedinthemicrowellsallowingEGFRantigentobecaptured.Theplateisthenwashedtoremoveanyunboundnon-specificmaterial.Aspecificrabbitanti-phospho-EGFRantibodyisaddedtodetectthecapturedphosphorylaterEGFR(Tyr1068)ontheplatewell.TheunbounddetectionantibodyiswashedawayfollowedbyincubationwithanHRP-conjugatedanti-rabbitantibody.AftertheadditionofTMBsubstrateandstopsolutiontheabsorbanceismeasuredat450nmusingaplatereader.Thisallowsforasensitiveenzymaticdetectionofthesample. Theentireassaytakeslessthan5hourstocompletewithminimalhands-ontime.Manyofthereagentsaresuppliedinready-touseformulationsforeaseofuse.ThekitalsoincludesastandardthatisrunasbothapositivecontrolandtogenerateastandardcurveforphosphorylatedEGFR(Tyr1068)measurement. II.EGFRBACKGROUND Epidermalgrowthfactorreceptor(EGFR)isareceptortyrosinekinasethatplaysakeyroleintheregulationofessentialnormalcellularprocessesandinthepathophysiologyofhyperproliferativediseasessuchascancer.EGFRisareceptortyrosinekinaseknowntobeessentialformediationofbothproliferativeandsurvivalsignalstocells.ActivationoftheEGFRsignalingpathwayhasbeenlinkedwithincreasedcellproliferation,angiogenesis,metastasis,anddecreasedapoptosis.EGFRisfoundinmostsolidtumorsanditsautophosphorylationactivatestheintrinsickinaseactivitytowardheterologoussubstrates,aswellascreatingdockingsitesforadapterproteinsformultipleadapterproteinstobindandnucleatesignalingcomplexesthatactivatetheRas,PI3Kinase,andPLCpathways.ManyoftheEGFRmutationsaredirectlyrelatedtovariousdiseasesandarethetargetofnumerousdrugs.Forinstance,theaminoacidsubstitutionL858RisoneofseveralheterozygousmutationsthathavebeenidentifiedinNon-Small-CellLungCancer(NSCLC)patientswhohaveclinicalresponsestotheEGFRinhibitorIressa?.ThereissomeevidencethatthesemutationsresultinelevatedactivityandenhancedsensitivitytoIressa?.InpatientswithtumorsbearingIressa?-sensitivemutations,resistantsubclonescontaininganadditionalEGFRmutation,T790M,emergeinthepresenceofthedrug.IthasbeenshownexperimentallythattheT790Mmutationleadstohigh-levelfunctionalresistancetoIressa?.InpatientswithtumorsbearingIressa?-sensitivemutations(eg.L858R,L861Q),resistantsubclonescontainingtheT790Mmutationemergeinthepresenceofthedrug.
MaterialsRequiredbutNotDelivered	1.Multi-channelorrepeatingPipettes 2.Plateshaker(optional) 3.Pipettorsandtipscapableofaccuratelymeasuring1-1000μL 4.GraduatedSEROlogicalpipettes 5.96-wellmicrotiterPlateReaderwith450nmfilter 6.Graphingsoftwareforplottingdataorgraphpaperformanualplottingofdata 7.Microfugetubesforstandardandsampledilutions 8.Mechanicalvortex 9.1litercontainer 10.Distilledordeionizedwater

ProductInformation
Components	CapturePlatepre-coatedwithanti-humanEGFRantibody:(PartNo.17-462A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch. Anti-EGFR(Tyr1068)detectionantibody:(PartNo.17-462B)Onebottle(11mL)ofanti-EGFR(Tyr1068)detectionantibodycontainingsodiumazide,readytouse. ELISADiluent:(PartNo.17-462C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse. 25XELISAWashBuffer:(PartNo.17-462D)Onebottle(50mL)of25XELISAWashBuffer. Anti-RabbitIgGHRPconjugate:(PartNo.17-462E)Onevial(125?L)of100Xanti-rabbitHRPconjugate. HRPDiluent:(PartNo.17-462F)Onebottle(25mL)ofHRPDiluent. TMBSolution:(PartNo.17-462G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse. StopSolution:(PartNo.17-462H)Onebottle(25mL)ofstopsolution,readytouse. PhosphorylatedEGFR(Tyr1068)Standard:(PartNo.17-462I)TwovialsofphosphorylatedEGFR(Tyr1068)standard,lyophilized. PlateCovers:Twoplatecovers.
Detectionmethod	Chromogenic

ProductInformation

Components

CapturePlatepre-coatedwithanti-humanEGFRantibody:(PartNo.17-462A)Onepre-coated96-stripwellimmunoplatesealedinafoilpouch.
Anti-EGFR(Tyr1068)detectionantibody:(PartNo.17-462B)Onebottle(11mL)ofanti-EGFR(Tyr1068)detectionantibodycontainingsodiumazide,readytouse.
ELISADiluent:(PartNo.17-462C)Onebottle(25mL)ofELISADiluentcontainingsodiumazide,readytouse.
25XELISAWashBuffer:(PartNo.17-462D)Onebottle(50mL)of25XELISAWashBuffer.
Anti-RabbitIgGHRPconjugate:(PartNo.17-462E)Onevial(125?L)of100Xanti-rabbitHRPconjugate.
HRPDiluent:(PartNo.17-462F)Onebottle(25mL)ofHRPDiluent.
TMBSolution:(PartNo.17-462G)Onebottle(25mL)ofstABIlizedtetramethylbenzidine(TMB),readytouse.
StopSolution:(PartNo.17-462H)Onebottle(25mL)ofstopsolution,readytouse.
PhosphorylatedEGFR(Tyr1068)Standard:(PartNo.17-462I)TwovialsofphosphorylatedEGFR(Tyr1068)standard,lyophilized.
PlateCovers:Twoplatecovers.

Detectionmethod

Chromogenic

StorageandShippingInformation
StorageConditions	Maintaintheunopenedkitat2-8°Cuntilexpirationdate.

Applications
Application	ThisPhospho-EGFR(Tyr1068)STARAssayELISAKitisusedtomeasure&quantifyEGFRlevels.
KeyApplications	ELISA

BIOLOGicalInformation
SpeciesReactivity	Human
AnalytesAvailable	EGFR
EntrezGeneNumber	NM_005228.3 NM_201282.1 NM_201283.1 NM_201284.1
GeneSymbol	EGFR ERBB1 ERBB mENA
Modifications	Phosphorylation
UniProtNumber	P00533
UniProtSummary	FUNCTION:SwissProt:P00533#Isoform2/truncatedisoformmayactasanantagoNIST. SIZE:1210aminoacids;134277Da SUBUNIT:BindsRIPK1.CBLinteractswiththeautophosphorylatedC-terminaltailoftheEGFreceptor.PartofacomplexwithERBB2andeitherPIK3C2AorPIK3C2B.TheautophosphorylatedforminteractswithPIK3C2B,maybeindirectly.InteractswithPELP1. SUBCELLULARLOCATION:Cellmembrane;Single-passtypeImembraneprotein.&Isoform2:Secreted. TISSUESPECIFICITY:Expressedinplacenta.Isoform2isalsoexpressedinovariancancers. PTM:PhosphorylationofSer-695ispartialandoccursonlyifThr-693isphosphorylated.&MonoubiquitinatedandpolyubiquitinateduponEGFstimulation;whichdoesnotaffecttyrosinekinaseactivityorsignalingcapacitybutmayplayaroleinlysosomaltargeting.Polyubiquitinlinkageismainlythrough"Lys-63",butlinkagethrough"Lys-48","Lys-11"and"Lys-29"alsooccur.DISEASE:SwissProt:P00533#DefectsinEGFRareassociatedwithlungcancer[MIM:211980]. SIMILARITY:SwissProt:P00533##Belongstotheproteinkinasesuperfamily.Tyrproteinkinasefamily.EGFreceptorsubfamily.&Contains1proteinkinasedomain. MISCELLANEOUS:BindingofEGFtothereceptorleadstodimerization,internalizationoftheEGF-receptorcomplex,inductionofthetyrosinekinaseactivity,stimulationofcellDNAsynthesis,andcellproliferation.

PhysicochemicalInformation
Sensitivity	Sensitivity:1unit/mL RangeofDetection:1.6to100units/mL

Dimensions

MaterialsInformation

MaterialsInformation

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Millipore/17-462 | Phospho-EGFR (Tyr1068) STAR Assay ELISA Kit/17-462/96 assays

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