
Description:
ProductName:MouseADIponectinELISA
#ofSamples:1X96Assays
IntendedUse: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:MouseAdiponectinELISA
Function: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:
Name | MouseAdiponectinELISA |
---|---|
RelatedProductNames | MouseAdiponectinELISAKitELISAKit;Adipocyte,C1qandcollagendomain-containingprotein;30kDaadipocytecomplement-relatedprotein;Adipocytecomplement-related30kDaprotein;ACRP30;Adiposemostabundantgenetranscript1protein;apM-1;Gelatin |
NCBIAccNumber | NP_004788.1 |
MolecularWeight | 26414 |
SwissProtNumber | Q15848 |
IntendedUse | ResearchUseOnly |