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 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