
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:
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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