
Description:
Vascularendothelialgrowthfactor(VEGF)isoneofthemostpotentandspecifictumor-angiogenicfactors.VEGFisusedbyoxygen-hungrycellstopromotegrowthofbloodvessels.Itbindstospecializedreceptorsonthesurfacesofendothelialcellsandinducestobuildnewvessels.MosttumorsproduceVEGFandinhibitionofVEGF-inducedangiogenesissignificantlyinhibitstumorgrowthinvivo.Anumberofantiangiogenicdrugshaveshowntobecapableofdecreasingthenumberofvessels. VEGFexpressioncorrelateswithmicrovesseldensityinanumberofsolidmalignanciesincludingcarcinomasofthebreast,lung,prostate,andcolon.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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