Description:Neutrophils,Mouse,mAbNIMP-R14,functionalstudyantibody,lowendotoxinThemonoclonalantibodyNIMP-R14ishighlyspecificformurineLy-6GandLy-6C.TheLy-6G/-6ClocusencodesafamilyofLy-6proteinsincludingLy-6GandLy-6C.Ly-6antigenshaveamolecularweightbetween15,000and18,000.Ly6GistogetherwithLy6cacomponentofthemyeloiddifferentiationantigenGr-1.Ly6GaGPI-anchoredproteinandisagoodMarkerofperipheralneutrophils.Althoughpredominantlypresentsonneutrophils,itisalsoexpressedonasubsetofeosinophils,differentiatingpre-monocytesandplasmacytoiddendriticcells.Ly6Cisamonocyte/macrophageandendothelialcelldifferentiationantigenregulatedbyinterferongamma,andmayplayaroleinthedevelopmentandmaturationoflymphocytes.Itisexpressedonbonemarrowcells,monocytes/macrophages,neutrophils,endothelialcells,andTcellsubsets.ExpressionofGr-1inbonemarrowcorrelateswithgranulocytedifferentiationandmaturation.However,thephysiologicalroleofLy6Galoneremainsstillunclear.ThemonoclonalantibodyNIMP-R14hasbeensuccessfullyusedtostainpolymorphonuclear(PMN)cellsandmonocytesforfluorescentactivatedcellsortingandinfrozenandparaffinsections.Treatmentwithantibodiesinvivoleadstoneutropeniaandhasinhibitoryeffectonlocalimmuneresponses.FurThermore,ithasbeenshowntobeusefulfordepletionofneutrophilsinmice.Itdepletesneutrophilsassoonas6hoursafterinjectionandupto6days.Specifications:CatalognumberHM1039-FSProducttypeMonoclonalantibodiesQuantity0.5mgFormulation0.5mgof0.2μmfilteredproteinGpurifiedantibodysolutioninPBSwithaconcentrationofatleast0.5mg/ml(exactconcentrationisindicatedonthelabel).Theendotoxinconcentrationis<24EU/mg,determinedwithHIT302LALAssay.ApplicationFlowcytometry,Frozensections,Functionalstudies,Immunoassays,ParaffinsectionsApplicationNotesIHC-F:Tissuewasfixedwithacetone(Ref6)IHC-P:Blockingwith20%normalrabbitserum(Ref4)FC:5x105cellswereincubatedwith10μg/mlantibody(Ref3)FS:Neutrophildepletion.MiceweretreatedwithNIMP-R14givenintraperitoneallyatadoseof1mg,6hbeforeinfection(Ref5)UseDilutionstobeuseddependondetectionsystemapplied.Itisrecommendedthatuserstestthereagentanddeterminetheirownoptimaldilutions.Thetypicalstartingworkingdilutionis1:50.ImmunogenPurifiedBALB/cmouseneutrophilsIsotypeRatIgG2bSpeciesMouseAliasLymphocyteantigen6complexlocusproteinG6c,myeloiddifferentiationantigenGr1Genename:Ly6gPositiveControlMouseneutrophilsNegativeControlMousethymocytesReferences1.Lopez,Aetal;Differentiationantigensonmouseeosinophilsandneutrophilsidentifiedbymonoclonalantibodies,BritJHaematology1984,57:4892.McLaren,Detal;Schistosomamansoni:evidencethatimmunityinvaccinatedandchronicallyinfectedCBA/Camiceissensitivetotreatmentwithamonoclonalantibodythatdepletescutaneouseffectorcells.ParasiteImmunol1987,9:6673.VanLent,Petal;Monocytes/macrophagesratherthanPMNareinvolvedinearlycartilagedegradationincationicimmunecomplexarthrititsinmice.JLeukocBiol1997,61:2674.Lubberts,Eetal;Adenoviralvector-mediatedoverexpressionofIL-4inthekneejointofmicewithcollagen-inducedarthritispreventscartilagedestruction.JImmunol1999,163:45465.Tacchini-Cottier,Fetal;AnimmunomodulatoryfunctionforneutrophilsduringtheinductionofaCD4+Th4responseinBALB/cmiceinfectedwithLeishmaniamajor.JImmunol2000,165:6286.Vriesde,Betal;ComplementfactorC5amediatesrenalischemia-reperfusioninjuryindependentfromneutrophils.JImmunol2003,170:38837.Nagendra,Setal;Absenceofcross-reactivitybetweenmurineLy-6CandLy-6G.CytometryA2004,58:1958.Navarini,Aetal;InnateImmune-induceddepletionofbonemarrowneutrophilsaggravatessystemicbacterialinfections.PNAS2009,106:71079.Paust,Hetal;Chemokinesplayacriticalroleinthecross-regulationofTh1andTh17immuneresponsesinmurinecrescenticglomerulonephritis.KidneyInternational,2012.10.Gao,Netal;MatrixMetalloproteinase-13asaTargetforSuppressingCornealUlcerationCausedbyPseudomonasaeruginosaInfection.JInfectDis.2015212:116StorageandstABIlityProductshouldbestoredat4 °C.Underrecommendedstorageconditions,productisstableforoneyear.PrecautionsForresearchuseonly.Notforuseinoronhumansoranimalsorfordiagnostics.Itistheresponsibilityoftheusertocomplywithalllocal/stateandFederalrulesintheuseofthisproduct.HycultBiotechisnotresponsIBLeforanypatentinfringementsthatmightresultwiththeuseoforderivationofthisproduct.DiseaseAutoimmunity,Infectiousdiseases