Description
CatalogueNumber	ECM300
BrandFamily	Chemicon®
TradeName	Quantimatrix Chemicon
Description	QuantiMatrixHumanFibronectinELISA
Overview	TheQuantiMatrix™ELISAkitforthedeterminationofsolublehumanfibronectinprovidessensitivitydownto10-20ng/mLasdeterminedfromculturesupernatantsandpurifiedproteinsamples. TheassayisintheformatofacompetitiveinhibitionELISA.Humanfibronectinispre-coatedonthewellswiththeexceptionofrowH,whichservesasareference.Standardsandsamplesaredilutedandpre-incubatedwith polyclonalrabbitantibodytohumanfibronectin.Thepolyclonalantibodybindstofibronectininthestandarddilutions,andinthesample,ifpresent.Themixtureisthentransferredtothehumanfibronectin-coatedplate.Freerabbitanti-humanfibronectinbindstothefibronectinontheplate.Goatanti-rabbitIgGHRPconjugatereactswithboundrabbitanti-fibronectin.WhenHRPsubstrateisadded,abluecolordevelops.Thereactionisstoppedbytheadditionofanacid,changingthecolortoyellow.ThiscolorisquantitatedusinganELISAreader;theintensityofthecolorisinverselyproportionaltotheamountoffibronectinintheoriginalsample.Astandardcurveisconstructedandsamplevaluesareinterpolated.
BackgroundInformation	ThedeterminationoflevelsofextracellularmatrixmoleculesisbecomingmoreimportanttounderstandingalterationsinmatrixfunctionduringdevelopmentandtumOrigenesis.Questionsofmatrixgeneregulationandfunctionrequireasensitivemeansofdeterminingthelevelsofproteinexpressionbycells.TheQuantiMatrix™ELISAkitforthedeterminationofsolublehumanfibronectinprovidessensitivitydownto10-20ng/mLasdeterminedfromculturesupernatantsandpurifiedproteinsamples. TheassayisintheformatofacompetitiveinhibitionELISA.Humanfibronectinispre-coatedonthewellswiththeexceptionofrowH,whichservesasareference.Standardsandsamplesaredilutedandpre-incubatedwithpolyclonalrabbitantibodytohumanfibronectin.Thepolyclonalantibodybindstofibronectininthestandarddilutions,andinthesample,ifpresent.Themixtureisthentransferredtothehumanfibronectin-coatedplate.Freerabbitanti-humanfibronectinbindstothefibronectinontheplate.Goatanti-rabbitIgGHRPconjugatereactswithboundrabbitanti-fibronectin.WhenHRPsubstrateisadded,abluecolordevelops.Thereactionisstoppedbytheadditionofanacid,changingthecolortoyellow.ThiscolorisquantitatedusinganELISAreader;theintensityofthecolorisinverselyproportionaltotheamountoffibronectinintheoriginalsample.Astandardcurveisconstructedandsamplevaluesareinterpolated.

ProductInformation
Components	QuantiMatrixTMFibronectinCoatedStrips(PartNo.60103)-1x96-wellstripplate HumanFibronectinStandard(PartNo.60109)-1lyophilizedbottle;reconstituteaccordingtotheprotocolonpage2 SampleDiluent(PartNo.60095)-1x15mLbottleof1%BSAinPBS+0.01%Thimerosal Goatanti-RabbitIgG-HRPConjugate(QM2)(PartNo.60110)-1bottle;readytouse Rabbitanti-HumanFibronectin(PartNo.60111)-1bottlepre-dilutedpolyclonalantibody;readytouse TMB/ESubstrate(PartNo.60096)-1bottle;readytouse StopSolution(PartNo.60193)-1bottle;(0.5MHClsolution) WashBufferConcentrate(PartNo.60245)-1x100mLbottle;providesfinalconcentrationsof0.05%Tween-20and0.01%Thimerosalwhenaddedto 900mLofdeionizedwaterasdescribedinstep1ofprotocolonpage2.
Detectionmethod	Chromogenic

StorageandShippingInformation

StorageConditions

Storethekitandallcomponentsat2-8°C. PRECAUTIONS: HumanfibronectinwaspurifiedfromhumanplasmathattestednegativeforHBsAgandHIV.However,allmaterialsshouldbehandledcarefullyandinaccordancewithgoodlaboratorypractices. Theinstructionsprovidedhavebeendesignedtooptimizeperformance.Deviationsfromthesuggestedproceduremayresultinsub-optimalperformanceofthekitandfailuretoproduceaccuratedata. Donotmixorsubstitutereagentsfromdifferentkitlots.

Applications
Application	TheQuantiMatrixELISAkitforthedeterminationofsolublehumanfibronectinprovidessensitivitydownto10-20ng/mLasdeterminedfromculturesupernatants&purifiedproteinsamples.
KeyApplications	ELISA

BIOLOGicalInformation
SpeciesReactivity	Human
AnalytesAvailable	Fibronectin
EntrezGeneNumber	NM_002026.2 NM_054034.2 NM_212474.1 NM_212475.1 NM_212476.1 NM_212478.1 NM_212482.1
EntrezGeneSummary	Thisgeneencodesfibronectin,aglycoproteinpresentinasolubledimericforminplasma,andinadimericormultimericformatthecellsurfaceandinextracellularmatrix.Fibronectinisinvolvedincelladhesionandmigrationprocessesincludingembryogenesis,woundhealing,bloodcoagulation,hostdefense,andmetastasis.Thegenehasthreeregionssubjecttoalternativesplicing,withthepotentialtoproduce20differenttranscriptvariants.However,thefull-lengthnatureofsomevariantshasnotbeendetermined.
GeneSymbol	FN1 DKFZp686O13149 DKFZp686H0342 DKFZp686I1370 MSF FN CIG DKFZp686F10164 LETS FINC
UniProtNumber	P02751
UniProtSummary	FUNCTION:SwissProt:P02751#Fibronectinsbindcellsurfacesandvariouscompoundsincludingcollagen,fibrin,heparin,DNA,andactin.Fibronectinsareinvolvedincelladhesion,cellmotility,opsonization,woundhealing,andmaintenanceofcellshape.InteractionwithTNRmediatesinhibitionofcelladhesionandneuriteoutgrowth(Bysimilarity). SIZE:2386aminoacids;262607Da SUBUNIT:Mostlyheterodimersormultimersofalternativelysplicedvariants,connectedby2disulfidebondsnearthecarboxylends;toalesserextenthomodimers.InteractswithFBLN1,AMBP,TNR,LGALS3BPandCOL13A1. SUBCELLULARLOCATION:Secreted,extracellularspace,extracellularmatrix. TISSUESPECIFICITY:PlasmaFN(solubledimericform)issecretedbyhepatocytes.CellularFN(dimericorcross-linkedmultimericforms),madebyfibroblasts,epithelialandothercelltypes,isdepositedasfibrilsintheextracellularmatrix.Ugl-Y1,Ugl-Y2andUgl-Y3arefoundinurine.DEVELOPMENTALSTAGE:Ugl-Y1,Ugl-Y2andUgl-Y3arepresentintheurinefrom0to17yearsofage. PTM:Sulfated. SIMILARITY:SwissProt:P02751##Contains12fibronectintype-Idomains.&Contains2fibronectintype-IIdomains.&Contains16fibronectintype-IIIdomains.

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