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Millipore/17-497 | Ras Activation ELISA ASSAY Kit/17-497/96 assays

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貨號: 17-497

品牌: Millipore

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Description
CatalogueNumber	17-497
Replaces	17-424
BrandFamily	Upstate
TradeName	Upstate
Description	RasActivationELISAASSAYKit
BackgroundInformation	RasproteinsfunctionasGDP/GTP-regulatedbinaryswitchesinsignaltransductioncascadesthatcanleadtocellgrowth,proliferation,differentiation,orsurvival.ThisRassuperfamilyhasatleastfivemajorbranchesthatincludeRas,Rho,Ran,Arf/Sar,Rab.Rasitselfhas3primaryisoforms(H-Ras,N-Ras,andK-Ras)thatdifferintheirapproximately20C-terminalaminoacids.TheyarecollectivelyreferredtoasRas.Initsactiveform,RasisboundtoGTP.Thiscausesaconformationalchangethatallowittointeractandbindtoseveraleffectormolecules,mostnotablythemembersoftheRaffamily,theRalGDSfamily,andPhosphoinositide3-kinases(PI3Kinase).RasthencleavesGTPtoGDPresultinginitsinactivation.Initsoncogenic,mutatedstate,RasisunabletohydrolyzeGTPtoGDP,thusstayinginanactivestateandactivatingnumerouspathways. Duetoitsroleasakeyregulatorofcellularfunctionsanditsimplicationsinvariouscancers,Rashasbeenapopulartargetforcancerresearchandanti-cancertherapeuticsforthepasttwodecadesforbothacademicandpharmaceuticalresearch.Itwasalsothefirsthumanoncogeneidentified.MutationsincellularRashavebeenfoundtobepresentinalargepercentageofallhumancancers.Morespecifically,K-Rasmutationsoccurfrequentlyinlung,pancreatic,andcoloncancers,whereasH-Rasmutationsareprevalentinbladder,kidney,andthyroidcancers,andN-Rasmutationsareassociatedwithmelanoma,hepatocellularcarcinoma,andleukemia(3).Inrecentstudies,RasactivityhasalsobeenimplicatedinanumberofhumandevelopmentdefectssuchasCostellosyndromeandNoonansyndrome(4). OnepaththatthepharmaceuticalindustryhastakentocontrolRasanditsactivityisbyattackingwhatsomeconsideritsAchilles’heel.Foritsactivation,Rasmustlocalizetotheplasmamembrane,butinterestingly,itlacksatransmembranedomain.Toachievethis,Rasmustfirstundergoapost-translationalmodificationknownasprenylationorgeranylationatitsC-terminalCAAXmotif.ThesedrugshaveyettopassclinicaltrialsthoughandthereisdoubtthattheywilleverbesuccessfulintreatingtumorsassociatedwithRasactivation.
MaterialsRequiredbutNotDelivered	1.Multi-channelorrepeatingPipettes 2.Plateshakeratroomtemperatureandat4oC. 3.Pipettors&tipscapableofaccuratelymeasuring1-1000μL 4.Graphingsoftwareforplottingdataorgraphpaperformanualplottingofdata 5.Multi-channelpipettorreservoirs 6.Proteaseinhibitors 7.Nucleasefreewater/deionizedwater 8.PBS 9.LuminometerorCCDcameracoupledimagingsystem

ProductInformation
Components	1.Glutathione-CoatedPlate,96-wellstripformat:(PartNo.3007063)1platecontaining12x8strip-wellscoatedwithglutathione. 2.Anti-RasDetectionAntibody:(PartNo.2006992)Onevialcontaining10μLofanti-Rasmousemonoclonalprimaryantibody.CanspecificallydetectK-,H-,N-isoformsofRas. 3.Mg2+Lysis/WashBuffer,5X:(PartNo.CS202617)1vialcontaining18mLof5XMg2+Lysis/WashBufferforpreparationofcelllysate. 4.20%Tween?20:(PartNo.#20-246)Onevialcontaining3mLof20%Tween?20. 5.10%BSAinTBS(BlockingBuffer):(PartNo.20-191)Onebottlecontaining10mL10%BSAin1XTBS. 6.20XTBSWash:(PartNo.20-190)Onebottlecontaining50mLof20XTBS,pH7.4. 7.GtxMs,HRP:(PartNo.CS202619)Oneambervialcontaining25μLofgoatanti-mouseHRPconjugatedantibody. 8.ChemiluminescentReactionBuffer:(PartNo.90496)Oneamberbottlecontaining8mLofChemiluminescentReactionBuffer.Keepoutoflight. 9.ChemiluminescentDetectionReagent:(PartNo.90495)Onebottlevialcontaining4mLofChemiluminescentDetectionReagent.Keepoutoflight. Storeat-80oC 10.Raf-1-RBD,GST:(PartNo.14-863)2vialscontaining150μLofRaf-1-RBDat2μg/μL. 11.EGFstimulatedHeLaCellLysate:(PartNo.12-500)2vialscontaining40μLoflysateat5mg/mLconcentration,lysedinMg2+Lysis/Washbuffer(Catalog#CS202617).

ProductInformation

Components

1.Glutathione-CoatedPlate,96-wellstripformat:(PartNo.3007063)1platecontaining12x8strip-wellscoatedwithglutathione.
2.Anti-RasDetectionAntibody:(PartNo.2006992)Onevialcontaining10μLofanti-Rasmousemonoclonalprimaryantibody.CanspecificallydetectK-,H-,N-isoformsofRas.
3.Mg2+Lysis/WashBuffer,5X:(PartNo.CS202617)1vialcontaining18mLof5XMg2+Lysis/WashBufferforpreparationofcelllysate.
4.20%Tween?20:(PartNo.#20-246)Onevialcontaining3mLof20%Tween?20.
5.10%BSAinTBS(BlockingBuffer):(PartNo.20-191)Onebottlecontaining10mL10%BSAin1XTBS.
6.20XTBSWash:(PartNo.20-190)Onebottlecontaining50mLof20XTBS,pH7.4.
7.GtxMs,HRP:(PartNo.CS202619)Oneambervialcontaining25μLofgoatanti-mouseHRPconjugatedantibody.
8.ChemiluminescentReactionBuffer:(PartNo.90496)Oneamberbottlecontaining8mLofChemiluminescentReactionBuffer.Keepoutoflight.
9.ChemiluminescentDetectionReagent:(PartNo.90495)Onebottlevialcontaining4mLofChemiluminescentDetectionReagent.Keepoutoflight.
Storeat-80oC
10.Raf-1-RBD,GST:(PartNo.14-863)2vialscontaining150μLofRaf-1-RBDat2μg/μL.
11.EGFstimulatedHeLaCellLysate:(PartNo.12-500)2vialscontaining40μLoflysateat5mg/mLconcentration,lysedinMg2+Lysis/Washbuffer(Catalog#CS202617).

StorageandShippingInformation

Applications
KeyApplications	ELISA CellFunctionAssay

BIOLOGicalInformation
SpeciesReactivity	Human Mouse Rat
AnalytesAvailable	Ras
EntrezGeneNumber	NP_203524.1 NP_001123914.1 NP_002515.1
EntrezGeneSummary	K-Ras Thisgene,aKirstenrasoncogenehomologfromthemammalianrasgenefamily,encodesaproteinthatisamemberofthesmallGTPasesuperfamily.AsingleaminoacidsubstitutionisresponsIBLeforanactivatingmutation.Thetransformingproteinthatresultsisimplicatedinvariousmalignancies,includinglungadenocarcinoma,mucinousadenoma,ductalcarcinomaofthepancreasandcolorectalcarcinoma.AlternativesplicingleadstovariantsencodingtwoisoformsthatdifferintheC-terminalregion.[providedbyRefSeq] H-Ras ThisgenebelongstotheRasoncogenefamily,whosemembersarerelatedtothetransforminggenesofmammaliansarcomaretroviruses.Theproductsencodedbythesegenesfunctioninsignaltransductionpathways.TheseproteinscanbindGTPandGDP,andtheyhaveintrinsicGTPaseactivity.Thisproteinundergoesacontinuouscycleofde-andre-palmitoylation,whichregulatesitsrapidexchangebetweentheplasmamembraneandtheGolgiapparatus.MutationsinthisgenecauseCostellosyndrome,adiseasecharacterizedbyincreasedgrowthattheprenatalstage,growthdeficiencyatthepostnatalstage,predispositiontotumorformation,mentalretardation,skinandmusculoskeletalabnormalities,distinctivefacialappearanceandcardiovascularabnormalities.Defectsinthisgeneareimplicatedinavarietyofcancers,includingbladdercancer,follicularthyroidcancer,andoralsquamouscellcarcinoma.Multipletranscriptvariants,whichencodedifferentisoforms,havebeenidentifiedforthisgene.[providedbyRefSeq]
GeneSymbol	KRAS K-Ras v-Ki-ras2Kirstenratsarcomaviraloncogenehomolog K-rasp21protein HRAS H-Ras Ha-Ras NRAS N-Ras neuroblastomaRASviral(v-ras)oncogenehomolog
UniProtNumber	P01116

PhysicochemicalInformation

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MaterialsInformation

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Millipore/17-497 | Ras Activation ELISA ASSAY Kit/17-497/96 assays

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