
Description:
Interleukin-6(IL-6)isamultifunctionalcytokinethatregulatestheimmuneresponse,hematopoiesis,theacutephaseresponse,andinflammation.DeregulationofIL-6productionisimplicatedinthepathologyofseveraldiseaseprocesses.Itslevelsareobservedinseveraldiseases,includingrheumatoidarthritis(RA).IL-6playsrolesinbothapro-inflammatoryandanti-inflammatorycytokine.ItissecretedbyTcellsandmacrophagestostimulateimmuneresponse.Inaddition,likeTNFa,IL-6isanotherADIpocytesecretoryproductthatmaybeinvolvedininsulinresistance.IL-6isacytokinesecretedbymanycells,includingadipocytesandadiposestromalcells.IL-6secretionisincreasedintheadipocytesofobesesubjectsandmaybeimportanteitherasacirculatinghormoneorasalocalregulatorofinsulinaction.IL-6hasbeenimplicatedinthedevelopmentofinsulinresistanceandtype2diabetesinobeseindividuals.LikeTNF,IL-6inhibitstheexpressionofLPL,but,unlikeTNF,IL-6doesnotstimulatelipolysis.Principle:
TheELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesaanti-ratcytokineantibodyforimmobilizationonthemicrotiterwellsandanti-ratcytokineantibodiesalongwithstreptavidinconjugatedtohorseradishperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwoantibodies,resultinginthemoleculesofthecytokinebeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.Theconcentrationofthecytokineisdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm.
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