Description
CatalogueNumber	CYT420
BrandFamily	Chemicon®
TradeName	ChemiKine Chemicon
Description	ChemiKine™PigmentEpitheliumDerivedFactor,SandwichELISA
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.
AlternateNames	PEDF
MaterialsRequiredbutNotDelivered	1.Multi-channelorrepeatingPipettes 2.37ºCincubator 3.Platesealers 4.Pipettors&tipscapableofaccuratelymeasuring10-1000mL 5.GraduatedSEROlogicalpipettes 6.96-wellmicroplatereaderwith450nmfilter 7.Graphpaperformanualplottingofdata 8.Polypropylenetesttubesforstandardandsampledilutions 9.Mechanicalvortex 10.One1or2litercontainer 11.Urea

ProductInformation
Components	PEDFELISAPlate:(PartNo.61597)One96-WellImmunoplatePre-coatedwithMouseanti-HumanPEDFMonoclonalAntibody,sealedinafoilpouch. WashBufferConcentrate:(PartNo.60245)One100mL(10X)bottleofConcentrate. AssayDiluent:(PartNo.60240)One60mLbottle(ReadytoUse). PEDFStandard(Human):(PartNo.61600)Onevialcontaining250ngrecombinantfulllengthhumanPEDF(Lyophilized). BiotinylatedMouseanti-HumanPEDFMonoclonalAntibody:(PartNo.61598)One50μLvial. Streptavidin,HRPconjugated:(PartNo.61599)One50μLvial. TMB/ESolution:(PartNo.60096)One10mLbottleofaReadytoUsesolutionof3,3",5,5"-tetramethylbenzidineinaproprietarybufferwithenhancer. StopSolution(PartNo.60193):One12mLbottleofanHClsolution(ReadytoUse).
Detectionmethod	Colorimetric

StorageandShippingInformation

StorageConditions

Maintaintheunopenedkitat2-8°Cuntilexpirationdateindicatedonthelabel.Afteropeningthekitmaintaineachcomponentat2-8°Cuntilexpirationdateindicatedonthelabel.Afterreconstitution,maintainthePEDFStandardat-20ºCforupto30days.Thestandardmaybefrozenandthawedthreetimeswithoutlossofimmunoreactivity.Donotstorereconstitutedstandardat4ºC,aslossofimmunoreactivitywillresult. Precautions ·WashBufferandAssayDiluentcontainthimerosal.Thimerosalishighlytoxicbyinhalation,contactwithskinorifswallowed.ThimerosalisapossIBLemutagenandshouldbehandledaccordingly. ·Theinstructionsprovidedhavebeendesignedtooptimizethekit"sperformance.Deviationfromtheinstructionsmayresultinsuboptimalperformanceofthekitandthefailuretoproduceaccuratedata.

Applications
Application	ThisChemiKinePigmentEpitheliumDerivedFactor,SandwichELISAisusedtomeasure&quantifyPEDFlevelsinNeuroscienceresearch.
KeyApplications	ELISA
ApplicationNotes	ChemiKine™PEDFkitisdesignedtomeasuretheamountofPEDFincellculturesupernatants,tissuehomogenatesandBIOLOGicalfluid(vitreous,serum,plasma,andserum-free)samplesofhumanorigin.Thereareenoughreagentsincludedinthiskitforone96-wellimmunoassayplate.Runningduplicatewellsforsamplesandstandardsisrecommended. ForResearchUseOnly.Notforuseindiagnosticprocedures.

BiologicalInformation
Host	Mouse
SpeciesReactivity	Human
AntibodyType	MonoclonalAntibody
AnalytesAvailable	PEDF
EntrezGeneNumber	NM_002615.4
GeneSymbol	SERPINF1 Serpin-F1 PEDF PIG35 EPC-1
UniProtNumber	P36955
UniProtSummary	FUNCTION:SwissProt:P36955#Neurotrophicprotein;inducesextensiveneuronaldifferentiationinretinoblastomacells.Potentinhibitorofangiogenesis.AsitdoesnotundergotheS(stressed)toR(relaxed)conformationaltransitioncharacteristicofactiveserpins,itexhibitsnoserineproteaseinhibitoryactivity. SIZE:418aminoacids;46342Da SUBCELLULARLOCATION:Secreted.Melanosome.Note=EnrichedinstageImelanosomes. TISSUESPECIFICITY:Retinalpigmentepithelialcellsandbloodplasma. DEVELOPMENTALSTAGE:Expressedinquiescentcells. DOMAIN:SwissProt:P36955TheN-terminal(AA44-121)exhibitsneuriteoutgrowth-inducingactivity.TheC-terminalexposedloop(AA382-418)isessentialforserpinactivity. PTM:TheN-terminusisblocked. SIMILARITY:Belongstotheserpinfamily.

PhysicochemicalInformation

Dimensions

MaterialsInformation

MaterialsInformation

Millipore 提供三种不同的 Immobilon PVDF 膜，是针对不同的蛋白质印迹应用的理想选择。 现在我们还 提供采用按规格裁切的印迹方片膜和两层过滤纸组成的印迹三明治。   与硝酸纤维素膜相比，Immobilon 膜具有多种优点。 它们不易破裂或卷 曲，便于裁剪且在切割过程中不会碎裂。 它们还有低背景、广泛溶剂相容性以及优异的着色能力。 此外，它们可以重复检测。 Immobilon-P 膜（沃比森-供应） ·         0.45μm 孔径 ·         建议用于大多数免疫印迹，特别在蛋 白质大于 20kDa 时 ·         Millipore 的快速免疫检测实验方法不需要膜封闭并缩短洗涤时间，将检测时间至少减少了两个小时，而不致降低灵敏 度或特异性 用于高通量实验室的印迹三明治 ·         当您需要处理多个印迹时，印迹三明治是一种便利省时的选择。 我们的印迹三明 治包含多片 Immobilon-P 膜，其中插入了按规格裁切的多张层析级印迹纸。 转印滤纸 ·         按照大多数免疫印迹尺寸进行裁切的层析级转印滤纸。 Immobilon-PSQ 膜（沃比森-供应） ·         0.2μm 孔径和大的内表面面积 ·         较其它膜具有更高的 蛋白质吸附率和测序得率 ·         建议用于小于 20kDa 的印迹蛋白质 ·         防止小分子量的蛋白质的渗漏 Immobilon-FL 膜（沃比森-供应） ·        针对荧光应用进行优化的转印膜 ·         极低的背景提高了所 有荧光检测实验的灵敏度 ·         与所有常用的荧光探针（所有激发和发射波长）兼容 ·         是多态性检测和化学荧光应用的理想选择