Description:
Granulocytecolony-stimulatingfactor(G-CSF)isacolony-stimulatingfactorhormone.Itisproducedbyanumberofdifferenttissuestostimulatethebonemarrowtoproducegranulocytesandstemcellsandtostimulatethebonemarrowtoreleasethemintotheblood.Italsostimulatesthesurvival,proliferation,differentiation,andfunctionofneutrophilprecursorsandmatureneutrophils,aswellasmonocytes.G-CSFinfluencesmonocytefunctionsinananti-inflammatoryway.ThestimulationofmonocyteswithG-CSFresultsinanattenuationofLPS-inducedreleaseofIL-1,TNF-a,IL-12andIL-18.Principle:
TheELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesamouseanti-humancytokineantibodyforimmobilizationonthemicrotiterwellsandgoatanti-humancytokineantibodiesalongwithstreptavidinconjugatedtohorserADIshperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwoantibodies,resultinginthemoleculesofthecytokinebeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.Theconcentrationofthecytokineisdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm. |
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