Description
CatalogueNumber	ECM101
BrandFamily	Chemicon®
TradeName	Millicoat Chemicon
Description	Millicoat™HumanFibronectinCoatedStrips(96-Wells)
Overview	Celladhesionplaysamajorroleincellularcommunicationandregulation,andisoffundamentalimportanceinthedevelopmentandmaintenanceoftissues.Scientistsarecontinuallyexaminingtheadhesionandmigrationofmanydiversecelltypesonvariousextracellularmatrix(ECM)componentproteins.Millicoat™CellAdhesionStripsareprovidedas12x8-wellremovablestripsinaplateframeforconvenienceandflexibilityindesigningassays.ThewellsinrowsA-GhavebeencoatedwithHumanFibronectin.RowHofeachstripiscoatedwithBSAwhichservesasanegativeassaycontrol.Cellsareseededontothecoatedsubstrate.Subsequently,adherentcellsarefixedandstained.RelativeattachmentisdeterminedusingabsorbancereADIngs.
AlternateNames	Formerly undertheCytoMatrixTMbrandname.

ProductInformation
Detectionmethod	Chromogenic

StorageandShippingInformation
StorageConditions	Maybestoredat2-8°Cinthefoilpouchforatleast3months.Unusedstripsmaybeplacedbackinthepouchforstorage.Ensurethatthedesiccantremainsinthepouch,andthatthepouchissecurelyclosed.

Applications
Application	MillicoatCellAdhesionStripsareprovidedas12x8-wellremovablestripsina96-wellplateframeforconvenience&flexibilityindesigningassays.
KeyApplications	ActivityAssay
ApplicationNotes	PROCEDURE: NOTE:Optimalassaytimingandperformancemayvaryfordifferentcelllinesbutgenerallycanbeobtainedusingsubconfluentcellculturesintheassaydescribedbelow.Subconfluentculturescanbeachievedbysplittingcells1to2dayspriortoperformingtheassay. 1.Rehydratethestripswith200mLofPBSperwellforatleast15minutesatroomtemperature.RemovethePBSfromtherehydratedstrips. 2.PrepareasinglecellsUSPension,preferablyusinganon-enzymaticdissociationbuffer.Optimumcelldensitymaybedeterminedbytitrationofthecells.Acommonstartingrangeisbetween1x10E05to1x10E07cells/mL. 3.Add100mLofthedilutedcellsuspensiontoeachwell.Incubatetheplateat37°Cfor1hourinaCO2incubator.Gentlywashtheplate2-3timeswithPBScontainingCa2+/Mg2+(200mL/well). 4.Add100mLof0.2%crystalvioletin10%ethanoltoeachwell.Incubatefor5minutesatroomtemperature.Removethestainfromthewells.Gentlywashtheplate3timeswithPBS(300mL/well). 5.Add100mLofSolubilizationBuffer(A50/50mixtureof0.1MNaH2PO4,pH4.5and50%ethanol)toeachwell.Allowstripstoincubateandgentlyshakeatroomtemperatureuntilthecell-boundstainiscompletelysolubilized;approximately5minutes. 6.Determinetheabsorbanceat540-570nmonamicroplatereader.

BIOLOGicalInformation
SpeciesReactivity	Human
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.
StemCellType	HumanEmbryonicStemCells MouseEmbryonicStemCells MesenchymalStemCells NeuralStemCells HematopoieticStemCells EpithelialCells PancreaticStemCells InducedPluripotentStemCells

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