Description:
Insulin-likegrowthfactor-I(IGF-I)actsasanimportantmediatorbetweengrowthhormoneandgrowththroughoutfetalandchildhooddevelopment.MorecircumstantialevidenceindicatestheassociationofIGF-Itotheriskofcancer.HighconcentrationsofIGF-Ihasbeenshowntobeanincreasedriskofcolorectalcancerandbreastcancerinsomestudiesandlessconsistentlywithprostate,thyroid,andhaematologicalmalignancies.IGF-IisapotentmitogenandimportantstimulusforADIpocytedifferentiation.IGF-IcanreducehyperglycemiainpatientswithsevereinsulinresistancebydirecteffectsmediatedviatheIGF-Ireceptor.IGF-Iinfusionlowersinsulinandlipidlevelsinhealthyhumansandreducesplasmaleptinconcentrationsinrats,suggestingthatIGF-Imayreducethedegreeofinsulinresistanceintype2diabetes,obesity,andhyperlipidemia.Principle:
TheELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesamouseanti-humancytokineantibodyforimmobilizationonthemicrotiterwellsandgoatanti-humancytokineantibodiesalongwithstreptavidinconjugatedtohorseradishperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwoantibodies,resultinginthemoleculesofthecytokinebeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.Theconcentrationofthecytokineisdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm. |
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