Description:
Granulocyte-macrophagecolony-stimulatingfactor(GM-CSF)isacytokinesecretedbymacrophages,Tcells,mastcells,endothelialcellsandfibroblasts. Itfunctionsasawhitebloodcellgrowthfactor.GM-CSFstimulatesstemcellstoproducegranulocytes(neutrophils,eosinophils,andbasophils)andmonocytes. Monocytesexitthecirculationandmigrateintotissue,wheretheymatureintomacrophages. IthasrecentlyshowedthatGM-CSFplaysakeyroleininflammatoryandautoimmunediseases.Principle:
Mouse CytokineELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesrabbitanti-mouseantibodiesforimmobilizationonthemicrotiterwellsandrabbitanti-mouseantibodiesalongwithstreptavidinconjugatedtohorserADIshperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwoantibodies,resultinginthemoleculesbeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.The concentrationofVEGFisdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm.
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