Description:ProductName:MouseADIponectinELISA#ofSamples:1X96AssaysIntendedUse:TheMouseAdiponectinELISAisasandwichenzymeimmunoassayforthequantitativemeasurementofmouseAdiponectin/Acrp30proteininserumandtissueculturemedium.Itisintendedforresearchuseonly.Principleofthetest:Surfaceofwellsinmicrotitrationplateiscoatedwithmonoclonalanti-mouseAdiponectinspecificantibody.Standards,QualityControls(QC)anddilutedsamplesarePipettedintothewells.AnymouseAdiponectin/Acrp30presentiscapturedbyimmobilizedantibodyandunboundproteiniswashedawayafterthefirstincubationperiod.Then,ahorseradishperoxidase(HRP)conjugatedpolyclonalanti-mouseAdiponectinantibodyisaddedtothewellsandincubated.Followinganotherwashingstep,toremoveunboundantibody-HRPconjugate,asubstratesolution(H2O2andTMB)isaddedtothewells.Theenzymaticreactionyieldsablueproductthatturnsyellowwhenacidicstopsolutionisadded.Theintensityofthecolour,measuredspectrophotochemicallyat450nm,isdirectlyproportionaltotheamountofthemouseAdiponectinboundintheinitialstep.ConcentrationofthedilutedtestsamplesarethenreadoffthestandardcurvethatisconstructedbyplottingtheabsorbancevaluesagainsteachrespectivemouseAdiponectinstandardlevel,usingafour-parameterfunction.ThedilutionfactorneedstobetakenintoconsiderationwhencalculatingtheactualconcentrationofAdiponectin/Acrp30analyteinthetestsamples.SensitivityandSpecificity:TheantibodiesintheMouseAdiponectinELISAkitarehighlyspecificformouseAdiponectinprotein.Nocross-reactivitywithseraofotheranimalspecies(rat,hamster,rabbit,sheep,goat,cattle,swine,horse)hasbeenobserved.TheassayrecognizesnativeandrecombinantmouseAdiponectin.Nocross-reactivityhasbeenobservedformousecytokines:RELM-?,RELM-?,Leptin,Leptin-receptor,Resistin;aswellasforratLeptinandhumanAdiponectinat100ng/ml.Onlyfractionalcross-reactivity0.3%hasbeenmeasuredforratrecombinantadiponectinat100ng/ml.ProductName:MouseAdiponectinELISAFunction:Importantadipokineinvolvedinthecontroloffatmetabolismandinsulinsensitivity,withdirectanti-diabetic,anti-atherogenicandanti-inflammatoryactivities.StimulatesAMPKphosphorylationandactivationintheliverandtheskeletalmuscle,enhancingglucoseutilizationandfatty-acidcombustion.AntagonizesTNF-alphabynegativelyregulatingitsexpressioninvarioustissuessuchasliverandmacrophages,andalsobycounteractingitseffects.InhibitsendothelialNF-kappa-BsignalingthroughacAMP-dependentpathway.Mayplayaroleincellgrowth,angiogenesisandtissueremodelingbybindingandsequesteringvariousgrowthfactorswithdistinctbindingaffinities,dependingonthetypeofcomplex,LMW,MMWorHMW.Subunit:Homomultimer.Formstrimers,hexamersand12-to18-mers.Thetrimers(lowmolecularweightcomplexes/LMW)areassembledvianon-covalentinteractionsofthecollagen-likedomainsinatriplehelixandhydrophobicinteractionswithintheglobularC1qdomain.Severaltrimerscanassociatetoformdisulfide-linkedhexamers(middlemolecularweightcomplexes/MMW)andlargercomplexes(highermolecularweight/HMW).TheHMW-complexassemblymayrelyaditionnallyonlysinehydroxylationandglycosylation.LMW,MMWandHMWcomplexesbindtoHBEGF,MMWandHMWcomplexesbindtoPDGFB,andHMWcomplexbindstoFGF2.SubcellularLocation:Secreted.TissueSpecificity:Synthesizedexclusivelybyadipocytesandsecretedintoplasma.Domain:TheC1qdomainiscommonlycalledtheglobulardomain.Ptm:HydroxylatedLys-33wasnotidentifiedinPubMed:16497731,probablyduetopoorrepresentationoftheN-terminalpeptideinmassfingerprinting.Ptm:HMWcomplexesaremoreextensivelyglycosylatedthansmalleroligomers.Hydroxylationandglycosylationofthelysineresidueswithinthecollagene-likedomainofadiponectinseemtobecriticallyinvolvedinregulatingtheformationand/orsecretionofHMWcomplexesandconsequentlycontributetotheinsulin-sensitizingactivityofadiponectininhepatocytes(Bysimilarity).Ptm:O-linkedglycansconsistofGlc-Galdisaccharidesboundtotheoxygenatomofpost-translationallyaddedhydroxylgroups.Ptm:NotN-glycosylated.Disease:DefectsinADIPOQarethecauseofadiponectindeficiency[MIM:605441].Theresultisaverylowconcentrationofplasmaadiponectin.Decreasedadiponectinplasmalevelsareassociatedwithobesityinsulinresistance,anddiabetestype2.Pharmaceutical:Adiponectinmightbeusedinthetreatmentofdiabetestype2andinsulinresistance.Miscellaneous:VariantsArg-84andSer-90showimpairedformationofHMWcomplexeswhereasvariantsCys-112andThr-164showimpairedsecretionofadiponectininanyform.Miscellaneous:HMW-complexbloodcontentsarehigherinfemalesthaninmales,areincreasedinmalesbycastrationanddecreasedagainuponsubsequenttestosteronetreatment,whichblocksHMW-complexsecretion(Bysimilarity).Intype2diabeticpatients,boththeratiosofHMWtototaladiponectinandthedegreeofadiponectinglycosylationaresignificantlydecreasedascomparedwithhealthycontrols.Similarity:Contains1C1qdomain.Similarity:Contains1collagen-likedomain.AdditionalInformation:NameMouseAdiponectinELISARelatedProductNamesMouseAdiponectinELISAKitELISAKit;Adipocyte,C1qandcollagendomain-containingprotein;30kDaadipocytecomplement-relatedprotein;Adipocytecomplement-related30kDaprotein;ACRP30;Adiposemostabundantgenetranscript1protein;apM-1;GelatinNCBIAccNumberNP_004788.1MolecularWeight26414SwissProtNumberQ15848IntendedUseResearchUseOnly