Description:LipopolysaccharideCore,mAbWN1222-5,functionalstudyantibody,lowendotoxinThemousemonoclonalantibodycloneWN1222-5recognizesthecoreregionoflipopolysaccharide(LPS),whereasitlacksreactivitywithfreelipidAandRd2orsmallerLPS.LPSisaconstituentoftheoutermembraneofthecellwallofcertaintypesofGram-negativebacteria,suchasE.coli,Salmonella,Shigella,Pseudomonas,Neisseria,Haemophilus,andsomeotherlesserknownpathogens.TheendotoxinsofGram-negativebacteriaareLPSmoleculeswiththreedistinctdomainsreferredtoaslipidA,coreoligosaccharideandO-polysaccharide.ThelipidAandcoreoligosaccharidecomprisetheendotoxincoreandarerelativelyconservedamongdifferentGram-negativebacterialspecies.TheO-polysaccharidesshowwidestructuraldiversityandgiverisetotheO-specificityofdifferentstrainandspeciesSEROtypes.LipidAisthetoxicmoietyofendotoxin,andiscovalentlylinkedtocoreoligosaccharideinallLPS.Rough(R)bacterialacktheO-polysaccharideoftheirsmooth(S)strainbacteriacounterparts,anddifferentroughmutantbacteriahavebeenisolatedexpressingarangeofincompletecoreoligosaccharidestructuresrangingfromcompletecore(Ra)todeep-rough(Re)mutantsexpressingonlylipidAlinkedtoinner-coreKDOresidues.Duringlife,humansaswellasothervertebrates,areoftenexposedtoLPS,forinstancebyenterobacteria.Whilegrowing,enterobacteriareleasessmallamountsofendotoxins.Predominantpartoftheendotoxinsstayonthecellwalluntilthebacteriumdisintegrates.Endotoxinsareheatstable,soevenboilingtheinfectedmaterialfor30minuteswillnotdenatureit.WaterydiarrheaiscausedbyreleasedLPSthatinteractswiththedigestivetrackintestine.CellsandtissuecanbedestroyedbytheLPSresultingininflammation.FurThermore,LPSisacausalfactorofNEC(necrotizingenterocolitis),adisorderfoundmostlyinnewborninfants.IfLPSgainsentrytothebloodstream,itcanbindthehostcells,suchasmacrophages.LPSresponsesarecomplicatedandinvolvecomplexesofLPS-bindingprotein(LBP),solubleand/ormembraneboundCD14andTolllikereceptor4(TLR4).LBPisaserumproteinthatcatalyzesLPSrecognitionbyCD14.RecognitionofLPStriggersacascadeofadversesystemicresponsesandorganfailure(septicshock).Specifications:CatalognumberHM6011-FSProducttypeMonoclonalantibodiesQuantity0.5mgFormulation0.5mgof0.2μmfilteredproteinGpurifiedantibodysolutioninPBSwithaconcentrationofatleast0.5mg/ml(exactconcentrationisindicatedonthelabel).Theendotoxinconcentrationis<24EU/mg,determinedwithHIT302LALAssay.ApplicationFunctionalstudies,Immunoassays,Paraffinsections,WesternblotApplicationNotesW:Anon-reducedsampletreatmentandSDS-PAGEwasused(Ref.1and5).IHC-P:Antigenretrievalwasperformedbyheatingsectionsin1xDIVADecloakerreagent.AspositivecontrolgutorlymphnodeofSIV-infectedrhesusmacaqueswasusedandasnegativecontrolgutorlymphnodeofnon-SIV-infectedrhesusmacaques(Ref.6).UseDilutionstobeuseddependondetectionsystemapplied.Itisrecommendedthatuserstestthereagentanddeterminetheirownoptimaldilutions.Thetypicalstartingworkingdilutionis1:50.Immunogen10E8heat-killedbacteriaIsotypeMouseIgG2aSpeciesN/AAliasLPScorePositiveControlLPSE.coliNegativeControlNormallymphnode(non-infected)References1.DiPadova,FEetal;Identificationofwidelycross-reactiveandcross-protectiveanti-LPScoremonoclonalantibodies(Mabs).InfectImmun1993,61:38632.Bahrami,Setal;Monoclonalantibodytoendotoxinattenuateshemorrhage-inducedlunginjuryandmortalityinrats.CritCareMed1997,25:10303.Pollack,Metal;Dualeffectsoflipopolysaccharide(LPS)antibodiesoncellularuptakeofLPSandLPS-inducedproinflammatoryfunctions.JImmunol1997,151:35194.Müller-Loennies,Setal;Identificationofacross-reactiveepitopewidelypresentinlipopolysaccharidefromenterobacteriaandrecognizedbythecrossprotectivemonoclonalantibodyWN1222-5.JBiolChem2003,278:256185.Tsuneyoshi,Netal;ThefunctionalandstructuralpropertiesofMD-2requiredforlipopolysaccharidebindingareabsentinMD-1.JImmunol2005,174:3406.Estes,Jetal;Damagedintestinalepithelialintegritylinkedtomicrobialtranslocationinpathogenicsimianimmunodeficiencyvirusinfections.PLOSpathogens2010,6:e10010527.Wang,Aetal;Anti-LPSTestStripfortheDetectionofFoodContaminatedwithSalmonellaandE.coli.MicrobialBiochemTechnol2011,3:28.Barclay,Getal;MonoclonalAntibodiestoEndotoxinCoreSegregateintoFamiliesofSpecificityandCross-ReactivitytoLipopolysaccharidesinELISAbyCorrelationClusterAnalysis.9.Cosgrove,Cetal;EarlyandnonreversIBLedecreaseofCD161++/MAITcellsinHIVinfection.Blood2013,121:95110.Ruan,Xetal.InVitroO-AntigenLigaseAssay.MethodsMolBiol2013,1022:18511.Belcher,Jeta;HemetriggersTLR4signalingleADIngtoendothelialcellactivationandvaso-occlusioninmurinesicklecelldisease.Blood2014,123:377StorageandstABIlityProductshouldbestoredat4 °C.Underrecommendedstorageconditions,productisstableforatleastoneyear.TheexactexpirydateisindicatedonthelabelPrecautionsForresearchuseonly.Notforuseinoronhumansoranimalsorfordiagnostics.Itistheresponsibilityoftheusertocomplywithalllocal/stateandfederalrulesintheuseofthisproduct.HycultBiotechisnotresponsibleforanypatentinfringementsthatmightresultfromtheuseorderivationofthisproduct.DiseaseInfectiousdiseases