Description:
Eotaxin-3,nowalsocalledchemokine(C-Cmotif)ligand26(CCL26)ormacrophageinflammatoryprotein4-alpha(MIP-4-alpha),isacytokinebelongingtotheCCchemokinefamily.Itattractsandactivateseosinophils,basophils,andTh2typeTlymphocytes.Itisexpressedbyseveraltissuesincludingheart,lungandovary,andinendothelialcellsthathavebeenstimulatedwiththecytokineinterleukin4.Principle:
TheELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesamouseanti-humancytokineantibodyforimmobilizationonthemicrotiterwellsandgoatanti-humancytokineantibodiesalongwithstreptavidinconjugatedtohorserADIshperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwoantibodies,resultinginthemoleculesofthecytokinebeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.Theconcentrationofthecytokineisdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm. |
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