Description:
IL-1aandIL-1ßareprominentagoNISTsmediatinginflammatoryandimmunomodulatoryeffects.Bothareproducedbymacrophages,monocytesanddendriticcells.Theyareimportantpartoftheinflammatoryresponseagainstinfection.Theyincreasetheexpressionofadhesionfactorsonendothelialcellstoenabletransmigrationofleukocytestositesofinfection.IL-1ßproductioninperipheraltissuehasalsobeenassociatedwithhyperalgesia(increasedsensitivitytopain)associatedwithfever.Principle:
TheELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesaanti-ratcytokineantibodyforimmobilizationonthemicrotiterwellsandanti-ratcytokineantibodiesalongwithstreptavidinconjugatedtohorserADIshperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwoantibodies,resultinginthemoleculesofthecytokinebeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.Theconcentrationofthecytokineisdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm.
Data:
