Description:
Epidermalgrowthfactor(EGF)isastrongmitogenformanycells,suchasfibroblasts.EGFalsostimulatestheproliferationofembryoniccells.TheproliferationofsomecelllineshasbeenshowntobeinhibitedbyEGF.EGFactsasadifferentiationfactorforsomecelltypes.ToalimitedextentEGFalsoaugmentsangiogenesisbecauseitismitogenicforendothelialcells.ThemitogenicactivityofEGFforendothelialcellscanbepotentiatedbythrombin.EGFisastrongchemoattractantforfibroblastsandepithelialcells.EGFaloneandalsoincombinationwithothercytokinesisanimportantfactormediatingwoundhealingprocesses.Principle:
TheELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesamouseanti-humancytokineantibodyforimmobilizationonthemicrotiterwellsandgoatanti-humancytokineantibodiesalongwithstreptavidinconjugatedtohorserADIshperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwoantibodies,resultinginthemoleculesofthecytokinebeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.Theconcentrationofthecytokineisdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm. |
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