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 Immobilon -P转印膜   PVDF膜 ·适用于免疫印迹、斑点印迹和其他蛋白质印迹的最佳PVDF膜；·Western Blotting、吸附分析、氨基酸分析、N端蛋白质测序、Dot/Slot blotting、糖蛋白显色和脂多糖分析；·与他相容的染液/染色剂：考马斯亮蓝染色剂、酰胺黑、印度墨水、丽春红、胶体金、CPTS、甲苯胺蓝、透射显影和Sypro Ruby染料. 产品名称Immobilon-P说    明应用于>20KD蛋白质的Western, dot/slot blotting和其他印迹孔    径0.45um尺寸cm×cm26.5×375数    量1卷目录编号IPVH00010 ――――――――――――――――――――――――――――――――――――――― Immobilon - NC膜，混合纤维素酯（Millipore） Immobilon-NC HATF由混合纤维素酯构成，不含细胞生长期间影响细胞完整的表面活性剂。使用这些膜可从事菌落转移实验。Immobilon-NC HAHY由混合纤维素酯构成，但含有转印过程期间能改善可湿性便于操作的表面活性剂。膜的孔径是0.45μm。用于Southern blotting，Northern Blotting，Western blotting，Colony/plaque Lifts，Dot/slot blots，吸附分析，细胞培养.。 产品名称Immobilon - NC说    明应用于Southern杂交， Northern杂交，免疫印迹，结合实验孔    径0.45um尺寸cm×cm33×300数    量1卷目录编号HATF00010