Overview | Introduction
PigmentEpithelium-DerivedFactor(PEDF)wasfirstidentifiedasafactorsecretedbyprimaryretinalpigmentepithelial(RPE)cellsthatpromotesthesurvivalofneuronsinvitroandinvivo.MolecularcharacterizationofPEDFshowsittobea50kDamemberoftheserpinfamilyofproteaseinhibitors1.AsecondmodeofactionofPEDFwasidentifiedwiththedemonstrationthatPEDFinducesapoptosisofmicrovascularendothelialcells,apropertythatmaintainstheavascularnatureoftheretina2.Asanumberofophthalmologicdisordersarecharacterizedbydysregulationoftheretinalinnervationandmicrovasculature,PEDFservesasanimportantregulatorofhumaneyedisease3.Inaddition,PEDFappearstohavemultifunctionalantitumoractivityinaneuroblastomamodel,asSchwanncell-derivedPEDFinducesadifferentiated,lessmalignantphenotypeinneuroblastomacells,promotesfurthergrowthandsurvivalofSchwanncells,andinhibitsangiogenesis4.
ChemiKine™PigmentEpithelium-DerivedFactor(PEDF)kitisasandwichstripwell-typeenzymeimmunoassay(EIA),whichmeasuresPEDF.ThekitwillmeasurePEDFofvitreousfluidofhumanorigin.
TestPrinciple
WiththeChemiKine™PEDFassaysystem,amousemonoclonalantibodygeneratedagainsthumanPEDFiscoatedontoamicroplateandisusedtocapturePEDFfromasample.Asecond,biotinylatedPEDF-specificmousemonoclonalantibodydetectsthecapturedPEDF.Peroxidase-labeledstreptavidindetectsthebiotinylatedmousemonoclonalantibody.AfteradditionofthesubstrateandstopsolutiontheamountofPEDFisdetermined.ThestandardcurvedemonstratesadirectrelationshipbetweenOpticalDensity(OD)andPEDFconcentration:i.e.,thehighertheODthehigherthePEDFconcentrationinthesample. |