Description:
Interleukin-2(IL-2)isacytokinethatplaysrolesinrespondingtomicrobialinfection.ItisproducedprimarilybyactivatedTcells.IL-2mediatesitseffectsbybindingtoitsreceptorsexpressedbyTcells,Bcells,NKcells,andAPCs.TheinteractionbetweenIL-2andIL-2Rleadstostimulationofthegrowth,differentiationandsurvivalofantigen-selectedcytotoxicTcellsviatheactivationoftheexpressionofspecificgenes.IL-2isalsonecessaryduringTcelldevelopmentinthethymusforthematurationofauniquesubsetofTcells.Insystemiclupuserythematosus(SLE),IL-2productionbyTlymphocytesinvitroisimpaired.Principle:
TheELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesamouseanti-humancytokineantibodyforimmobilizationonthemicrotiterwellsandgoatanti-humancytokineantibodiesalongwithstreptavidinconjugatedtohorserADIshperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwoantibodies,resultinginthemoleculesofthecytokinebeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.Theconcentrationofthecytokineisdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm. |
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