
Description:
MacrophageInflammatoryProtein-1(MIP-1)isamemberoftheC-Csubfamilyofchemokinesthatexhibitavarietyofproinflammatoryactivitiesinvitroincludingleukocytechemotaxis. MIP-1isamajorfactorproducedbymacrophagesafterstimulatedwithbacterialendotoxins.Itactivatesgranulocytes(neutrophils,eosinophilsandbasophils)whichcanleadtoacuteneutrophilicinflammation.Italsoinducesthesynthesisandreleaseofotherpro-inflammatorycytokinessuchasIL-1,IL-6andTNFafromfibroblastsandmacrophages. Inadditiontoitsproinflammatoryactivities,MIP-1inhibitstheproliferationofhematopoieticstemcellsinvitroandinvivo.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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