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Millipore/15-107 | MAPK Pathway 1 Explorer Antibody MiniPack/15-107/3 vials/Pk

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貨號: 15-107

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Description
CatalogueNumber	15-107
BrandFamily	Upstate
TradeName	Upstate
Description	MAPKPathway1ExplorerAntibodyMiniPack
AlternateNames	OncogeneRAF1 rafproto-oncogeneserine/threonineproteinkinase
BackgroundInformation	PathwayExplorerAntibodyMiniPack: EachPathwayExplorerAntibodyMinipackcontainsthreerelatedantibodiesaspartofasignalingcascadeoracombinationoftotalandphosphorylatedformsofkeysignalingtargets.Eachofthethreeantibodiesare30%theoriginalpacksize.FullsizeversionsofeachofthePathwayExplorerantibodiesareavailableforsaleindividuallyunderthesamecatalognumberwiththeremovalof“SP”offofeachone(e.g.05-591SPcanbeorderedas05-591). Erk1/2: Erk(Extracellularsignal-RelatedKinase)isafamilyoftwo,highlyhomologousproteinsdenotedasErk1(p44,MAPK3)andErk2(p42,MAPK1)thatbothfunctioninthesamepathway.ThetwoproteinsareoftenreferredtocollectivelyasErk1/2orp44/p42MAPkinase.TheErkpathwayisconsideredtheclassical,canonicalMAPK(Mitogen-ActivatedProteinKinase)signalingpathway.Itisanevolutionarilyconservedpathwaythatcontrolsandisacriticalregulatorofthegrowthandsurvivalthroughthepromotionofcellproliferationandthepreventionofapoptosis.Erkisinvolvedinthecontrolofmanyfundamentalcellularprocessesincludingcellproliferation,survival,differentiation,apoptosis,motilityandmetabolism.Erkisactivatedbygrowthfactorstimulationofreceptortyrosinekinases(RTKs),GPCR,and/orintegrinstimulation.ThisactivatestheRas-Raf-MEK-Erkpathwaythatresultsinthephosphorylation/activationofErk1/2(p44/p42)ontheTxYmotif(Thr202/Tyr204andThr185/Tyr187forErk1&Erk2,respectively). Raf: TheRafproteins(A-Raf,B-Raf,Raf-1/c-Raf)areSer/ThrkinaseswithhomologytothePKCfamily,containinganN-terminalregulatorydomainandaC-terminalcatalyticdomain.Raf(MAPKinaseKinaseKinase)isthetopkinaseinthecanonicalMAPKpathway.MembersoftheRaffamilybindtoactivatedRas.Ras,initsGTP-boundactivestate,activatesthekinaseactivityofRaf.ThisresultsinRaftranslocationtotheplasmamembraneandactivation.Onceactivated,RafthenbindstoandactivatesMEK(MAPKinaseKinase)byphosphorylatingitonthetworesiduemotif.ItisthoughtthatB-RafmightbethepredominantactivatorofMEKandthatRaf-1hasaroleinprotectionagainstapoptosis;aprocessthatdoesnotrequireeitheritskinaseactivityoritsactivationofMEK.TheregulationofalargenumberofcellularprocessesaredependentupontheactivationstateofERK,sothecontrollingofthispathwaycouldhaveprofoundeffectsonvariousdiseases.ActivationofRaf-1involvesphosphorylationofSer338/339andTyr340/341.ActivatingmutationsofB-Rafthatdisruptitsauto-inhibitionloophavebeenimplicatedinanumberofcancers,includingmelanomaandcoloncancer. MEK1/MEK2: MAPKinase/ErkKinase(MEK),alternativelyknownasMKK,isatruedual-specificitykinase,inthatitphosphorylatestheMAPkinases(Erk1/2;p44/p42))onboththeThrandTyroftheactivationmotifTEY.Invitro,thetyrosinephosphorylationisfavored,whereasinvivobothphosphorylationeventsappeartooccursimultaneously.Thissuggeststhatanadditionalfactorispresentincellstofacilitatethereaction.MEK1andMEK2areactivatedbyphosphorylationoftwoserineresidues(Ser218/222inMEK1andSer222/226inMEK2),whicharesubstratesfortheRaffamilyofkinases.MutationofthephosphorylationsitesfromSertoAspcreatesaproteinwithconstitutivekinaseactivity,whichwhenexpressedincellsisabletocausetransformation. *Seefullsizeversionsforcorrespondingreferences.

ProductInformation
Components	05-538SPAnti-phospho-Raf-1(Ser338) 05-747SPAnti-phospho-MEK1(Ser218/222)/MEK2(Ser222/226) 05-797RSPAnti-phospho-Erk1/2 (Thr202/Tyr204,Thr185/Tyr187),recombinantcloneAW39R
Format	Purified
Control	05-538SP:3T3/NIHcells
Presentation	3individualtubescontainingeitherAnti-phospho-Raf-1(Ser338);orAnti-phospho-MEK1(Ser218/222)/MEK2(Ser222/226);Anti-phospho-Erk1/2 (Thr202/Tyr204,Thr185/Tyr187),recombinantcloneAW39R
Properties	Eachvialis30%thesizeoftheparentcatalognumber

StorageandShippingInformation
StorageConditions	Stablefor1yearat-20°Cfromdateofreceipt. HandlingRecommendations:Uponreceipt,andpriortoremovingthecap,centrifugethevialandgentlymixthesolution.Aliquotintomicrocentrifugetubesandstoreat-20°C.Avoidrepeatedfreeze/thawcycles,whichmaydamageIgGandaffectproductperformance.

Applications
Application	ThisAntibodypackcontainsAnti-phospho-Raf-1Antibody(Ser338),Anti-phospho-MEK1Antibody(Ser218/222)/MEK2(Ser222/226),Anti-phospho-Erk1/2Antibody(Thr202/Tyr204,Thr185/Tyr187),validatedforuseinIF,Flow,WB,Multiplexing
KeyApplications	Immunofluorescence FlowCytometry WesternBlotting Multiplexing
ApplicationNotes	05-797RSP:ImmunofluorescenceAnalysis:A431cellswerefixed,permABIlizedanddualstainedwithanti-phospho-Erk. FlowCytometryAnalysis:Jurkatcellswerefixed,permeablized,andstainedwithanti-phospho-Erk1/2(Thr202/Tyr204,Thr185/Tyr187).

BIOLOGicalInformation
Immunogen	05-797RSP:PhosphorylatedpeptideencompassingtheTxYmotifintheactivationloopofErk1andErk2.;05-747SP:KLH-conjugated,syntheticpeptidecontaining…DpSMANpSF…inwhichpScorrespondstophosphorylatedaminoacids218and222ofhumanMEK1.Theimmunizingsequenceishighlyconserved.;05-538SP:KLH-conjugated,syntheticpeptideencompassingandincludingphosphorylatedSer338ofhumanRaf-1.Theimmunizingsequenceisidenticalinratandchickenandhas12/13identicalaminoacidsinXenopus.
Epitope	Pleaseseerelevantdatasheet.
Clone	05-797RSP:AW39R
Concentration	05-538SP:1mg/mL
Specificity	05-797RSP:RecognizesErk1&2onlywhenduallyphosphorylatedonitsTxYactivationmotif.;05-747SP:PhosphorylatedMEK1/MEK2;05-538SP:RecognizesRaf-1phosphorylatedatSerine338
SpeciesReactivity	Human Mouse Rat Chicken Xenopus
SpeciesReactivityNote	05-538SP:Mouseandhuman. Predictedtocrossreactwithrat,chickenandXenopusbasedonsequencehomology.
EntrezGeneNumber	NP_203524 NP_002871 NP_002746 NP_109587 NP_002737 NP_002736
GeneSymbol	MAP2K1 PRKMK1 MKK1 MAPKK1 MEK1 RAF1 cRaf Raf-1 c-Raf RAF CRAF C-RAF MAPK3 ERK1 p44 P44ERK P44MAPK Erk2 MAPK1 P42MAPK P42ERK
Modifications	Phosphorylation
UniProtNumber	P04049 Q02750 P36507 P27361 P28482
UniProtSummary	Raf-1: FUNCTION:Involvedinthetransductionofmitogenicsignalsfromthecellmembranetothenucleus.PartoftheRas-dependentsignalingpathwayfromreceptorstothenucleus.ProtectscellsfromapoptosismediatedbySTK3. CATALYTICACTIVITY:ATP+aprotein=ADP+aphosphoprotein. COFACTOR:Binds2zincionspersubunit. SUBUNITSTRUCTURE:InteractswithRasproteins.TheinteractionwithRasisantagonizedbyRIN1.WeaklyinteractswithRIT1Bysimilarity.InteractswithSTK3,whichinhibitsitspro-apoptoticactivity. TISSUESPECIFICITY:Inskeletalmuscle,isoform1ismoreabundantthanisoform2. POST-TRANSLATIONALMODIFICATION:PhosphorylateduponDNAdamage,probablybyATMorATR.PhosphorylationatThr-269increasesitskinaseactivity. INVOLVEMENTINDISEASE:DefectsinRAF1arethecauseofNoonansyndrometype5(NS5)[MIM:611553].Noonansyndrome(NS)isadisordercharacterizedbydysmorphicfacialfeatures,shortstature,hypertelorism,cardiacanomalies,deafness,motordelay,andableedingdiathesis.Itisageneticallyheterogeneousandrelativelycommonsyndrome,withanestimatedincidenceof1in1000-2500livebirths.Ref.24Ref.25 DefectsinRAF1arethecauseofLEOPARDsyndrometype2(LEOPARDsyndrome-2)[MIM:611554].LEOPARDsyndromeisanautosomaldominantdisorderallelicwithNoonansyndrome.TheacronymLEOPARDstandsforlentigines,electrocardiographicconductionabnormalities,ocularhypertelorism,pulmonicstenosis,abnormalitiesofgenitalia,retardationofgrowth,anddeafness. MEK1: FUNCTION:CatalyzestheconcomitantphosphorylationofathreonineandatyrosineresidueinaThr-Glu-TyrsequencelocatedinMAPkinases.ActivatesERK1andERK2MAPkinases. CATALYTICACTIVITY:ATP+aprotein=ADP+aphosphoprotein. ENZYMEREGULATION:Activatedbyphosphorylation. SUBUNIT:InteractswithMORG1(Bysimilarity).InteractswithYersiniayopJ. PTM:PhosphorylationonSer/ThrbyMAPkinasekinasekinases(RAForMEKK1)regulatespositivelythekinaseactivity. PTM:AcetylationbyYersiniayopJpreventsphosphorylationandactivation,thusblockingtheMAPKsignalingpathway. DISEASE:DefectsinMAP2K1areacauseofcardiofaciocutaneoussyndrome(CFCsyndrome)[MIM:115150];alsoknownascardio-facio-cutaneoussyndrome.CFCsyndromeischaracterizedbyadistinctivefacialappearance,heartdefectsandmentalretardation.Heartdefectsincludepulmonicstenosis,atrialseptaldefectsandhypertrophiccardiomyopathy.Someaffectedindividualspresentwithectodermalabnormalitiessuchassparse,friablehair,hyperkeratoticskinlesionsandageneralizedichthyosis-likecondition.TypicalfacialfeaturesaresimilartoNoonansyndrome.Theyincludehighforeheadwithbitemporalconstriction,hypoplasticsupraorbitalridges,downslantingpalpebralfissures,adepressednasalbridge,andposteriorlyangulatedearswithprominenthelices.TheinheritanceofCFCsyndromeisautosomaldominant. SIMILARITY:Belongstotheproteinkinasesuperfamily.STESer/Thrproteinkinasefamily.MAPkinasekinasesubfamily. SIMILARITY:Contains1proteinkinasedomain. Erk2: FUNCTION:CatalyzestheconcomitantphosphorylationofathreonineandatyrosineresidueinaThr-Glu-TyrsequencelocatedinMAPkinases.ActivatesERK1andERK2MAPkinases. CATALYTICACTIVITY:ATP+aprotein=ADP+aphosphoprotein. ENZYMEREGULATION:ActivatedbyphosphorylationontyrosineandthreonineinresponsetoinsulinandNGF.Bothphosphorylationsarerequiredforactivity SUBUNITSTRUCTURE:InteractswithMORG1Bysimilarity.BindstoHIV-1NefthroughitsSH3domain.Thisinteractioninhibitsitstyrosine-kinaseactivity.InteractswithitssubstratesHSF4andARHGEF2.InteractswithNISCH. DOMAIN:TheTXYmotifcontainsthethreonineandtyrosineresidueswhosephosphorylationactivatestheMAPkinases. PTM:DuallyphosphorylatedonThr-185andTyr-187,whichactivatestheenzyme.
MolecularWeight	05-797RSP:42and44kDa;05-747SP:45kDa;05-538SP:74kDa

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