Signosis/Rat IL1β ELISA/EA3005/1 Ea

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: