Description:
Nervegrowthfactor(NGF)isasmallsecretedproteinwhichinducesthedifferentiationandsurvivalofparticulartargetneurons(nervecells).NGFhasrecentlybeenidentifiedasanovelangiogenicmolecule,whichexertsavarietyofeffectsinthecardiovascularsystemandonendothelialcells.NGFmaycontributetomaintenance,survival,andfunctionofendothelialcellsbyautocrineand/orparacrinemechanisms.Beta-NGF(β-NGF)isstructurallyrelatedtoBDNF,NT-3andNT-4.NGFisapotentneurotrophicfactorthatsignalsthroughitsreceptorβ-NGFR,andplaysacrucialroleinthedevelopmentandpreservationofthesensoryandsympatheticnervoussystems.β-NGFalsoactsasagrowthanddifferentiationfactorforBlymphocytesandenhancesBcellsurvival.Principle:
TheELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesamouseanti-humancytokineantibodyforimmobilizationonthemicrotiterwellsandgoatanti-humancytokineantibodiesalongwithstreptavidinconjugatedtohorserADIshperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwoantibodies,resultinginthemoleculesofthecytokinebeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.Theconcentrationofthecytokineisdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm. |
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