ProductDescription
PureCol®CollagenCoated,96-wellPlateswithaflatbottomhaveanuniformandconsistentapplicationofhighqualityTypeIcollagenonclearpolystyrenesurface.PlatesarecompatIBLeforusewithallcommondispensers,readersandwashers.Platesareasepticallyprocessedandarenon-pyrogenic.Platesareavailableinquantitiesof5platespersleeve.
| Parameter,Testing,andMethod | PureCol®Coated96-MultiwellPlate#5072 |
| SterilizationMethod | AsepticallyProcessed |
| Size | 96-MultiwellPlate |
| CollagenUsed | PureCol®TypeI |
| StorageTemperature | 2-30°C |
QuantityperPackage | 5Plates |
| ShelfLife | Minimumof6monthsfromdateofreceipt |
| FlaskPolymer | Polystyrene |
| TisueCultureTreatedPriortoCoating | Yes |
| GrowthAreaperWell | 0.35cm2 |
| NominalVolumeperWell | 320uL |
| TypicalWorkingVolumeperWell | 50-200uL |
ProductQ&A
WecompletedastudytoshowthatDNAiscompletelydestroyedatpH2,anddemonstratedthatourcollagenproductsdonotcontainDNA.
Thecollagenisfullyhydrolyzed.TheaminoacidanalysisisdoneusingtheWatersAccQ-Tagderivatizationmethod.Duringtheacidhydrolysisstep,asparagine(N)isconvertedtoasparticacid(D)andglutamine(Q)isconvertedtoglutamicacid(E).Tryptophan(W),ifpresent,isdestroyedduringacidhydrolysis.Experimentally,onecandeterminethepicomoles(pmol)ofeachaminoacidperinjecteddetectedusingaminoacidstandards.Fortheconcentrationdetermination,thetotalnumberofpmolofeachaminoacidissummedtogetthetotalpmolofthe18aminoacidsdetected.Thetotalpmolaminoacidsisdividedbythetheoreticalnumberofaminoacidresiduesincollagenbasedonthepublishedsequence.Theresultisthepmolofcollageninjected.Theresultisthenmultipliedbythedilutionand300,000isusedasthecollagenmolecularweighttogettomg/mL.Themolecularweightofcollagenisnotwellagreedupon.
Dilutingwith1XPBS(ratherthanwateror0.01NHCl)wouldhaveaneffectforcoatingpurposes.ItwouldchangethepHofthedilutedcollagensolutionfromacidtoneutralpH.ThepHchangewilltransformthecollagenmoleculesfromamolecularformtoafibrillarform;andthenthenatureofcoatingsurfacewillbechangedfromamonomericcoatingtoafibrillarcoating.
WeusethefollowingantibodiesfromSouthernBiotech:
1.1310-02–GoatAnti-TypeICollagen-FITC
2.1310-08–GoatAnti-TypeICollagen-BIOT
3.7100-05–Streptavidin-HRP
ThemajorcollagenmolecularspeciesinourTypeIcollagenproductsaremonomers(approx.70%),buttherearedimers,trimersandafewpercentagesofoligomerstoo(approx.30%)withsomeminoramountsofcollagenfragments.Thecollagenmonomerisarodshapedmoleculewith300nminlengthand1.5nmindiameter.Thedimer,trimerandoligomerare600nm,900nmandevenlongerinlengthrespectively.Accordingtothecoatingprocedures,thecollagenmoleculesareattachedtothechargedpolystyrenesurfacerandomlybychargeoraffinityinacidconditionsduringthe1-2hrsincubationperiodat37°C,andanyunattachedmaterialsareremovedbyaspirationandrinsing.Therefore,thecoatedsurfaceisasinglelayerofcollagenmonomer,dimer,trimerandoligomermixtures.Thethicknessofthemono-molecularlayerisdependentonhowthosemoleculesareattachedonthesurface.Thecoatingdensitythicknesswouldgenerallybecharacterizedasa1moleculethicknesswhichcouldberangingfromafewnanometerstoafewhundrednanometerswiththewholesurfacebeingcoveredbycollagen.
ThenetchargeofTypeIcollagenproducts’(PureCol®,BovineCollagenandVitroCol®,HumanCollagen)moleculeisdirectlyrelatedtothepH.AtanacidicpH,theaminoacids(zwitterions)alongthecollagenmoleculearepositivelycharged,makingtheentirecollagenmoleculepositive.Attheisoelectricpoint(orzone)ofcollagen,aroundpH7-8,theaminoacidsalongthecollagenmoleculearepositivelyandnegativelycharged,makingthenetchargeofthecollagenmoleculeclosetozero.AtabasicpH,theaminoacidsalongthecollagenmoleculewerenegativelycharged,makingtheentirecollagenmoleculenegative.
Further,thenatureofthechargeofthecollagencoatingsurfacewillbedependentonthetypeofcoatingapplied.ForamonomericcollagencoatingswhenthecollagenisappliedunderanacidicpHcondition,thesurfaceispositivelycharged.IfthesurfaceisrinsedwithpHneutralbufferormediathenitwillchangethechargeofthecollagensurfacenetchargeclosetozero.Fora3Dgelcoating,thecollagenpreparedunderneutralpH;thenetchargeofthecollagensurfaceisclosetozero.
Usingrotaryshadowingtechniqueundertransmissionelectronmicroscopy,itwasfoundthatourcollagen,onaverage,consistsofapproximately80%monomers,13%dimers,trimers,andoligomerswiththeremaining7%collagenfragments.
Yes.ThecollagenmoleculeinPureCol,Nutragen,VitroCol,andallofourotherAtelocollagenproductswerepreparedfromnativecollagenmatrixbypepsintreatmentundercontrolledconditionstoremovethenon-helicalportion,telo-peptides,onlyandthehelicalportionisintact.Inthiscase,theenzymaticactivesitesforMMP(MatrixMetalloproteinase),suchasforMammalianCollagenaseMatrixMetalloproteinase8(MMP-8),onthemoleculewaspreserved.
Thesepepsintreatedcollagenproductsshouldbehaveasnativeintactcollagen.
TGFbetawouldhavebeendigestedwiththepepsinenzymaticdigestionstep.ItwasundetectablebySDSPAGEsilverstainaswell.Wedidn’tdoanyspecificmeasurementsbyELISAhoweverbutpresencesofTGFbetaisnotanticipated.
WeprimarilyusetheBiuretmethod,butwealsouseBCA,AAA,andhydroxyl-prolineassays.
-Collagensolutionsthatarefrozentendtohaveissuesforming3Dhydrogels,andwilllikelynotwork.Thesolutionsshouldstillbegoodfor2Dcoatings.
-Collagensolutionsthatareleftoutatroomtemperatureforextendedperiodsoftimemayshowsignsofdegradation,whichwillaffecttheformationof3Dhydrogels.Itislikelystillfinefor2Dcoatings.
Ourrecommendationisthis:Ifyouareusingtheproductdirectlyforapublication,wehighlysuggestbuyinganewbottleiftheoneyouhavewascompromised.
ProductReferences
BecausePureCol®hasbeencitedinover2000publications,wehaveonlypostedafewbelow:
Sorensen,JacobR.,etal."Analteredresponseinmacrophagephenotypefollowingdamageinagedhumanskeletalmuscle:implicationsforskeletalmusclerepair."TheFASEBJournal(2019):fj-201900519R.
Sorensen,JacobR.,etal."Analteredresponseinmacrophagephenotypefollowingdamageinagedhumanskeletalmuscle:implicationsforskeletalmusclerepair."TheFASEBJournal(2019):fj-201900519R.
Colaço,E.,etal."HierarchicalCollagen-HydroxyapatiteNanostructuresDesignedThroughLayer-by-LayerAssemblyofCrystal-DecoratedFibrils."J.,HierarchicalCollagen-HydroxyapatiteNanostructuresDesignedThroughLayer-by-LayerAssemblyofCrystal-DecoratedFibrils(May13,2019)(2019).
Schwerdtfeger,LukeA.,etal."Humancolonfunctionexvivo:Dependenceonoxygenandsensitivitytoantibiotic."PloSone14.5(2019):e0217170.
Cardoso,Ana,etal."MiR-144overexpressionasapromisingtherapeuticstrategytoovercomeglioblastomacellinvasivenessandresistancetochemotherapy."Humanmoleculargenetics(2019).
Steele,HannahE.,etal."MechanotransductionofmitochondrialAMPKanditsdistinctroleinflow-inducedbreastcancercellmigration."Biochemicalandbiophysicalresearchcommunications514.2(2019):524-529.
Gehwolf,Renate,etal."GlobalResponsesofIl-1β-Primed3DTendonConstructstoTreatmentwithPulsedElectromagneticFields."Cells8.5(2019):399.
Alexander,Frank,SebastianEggert,andDoriellePrice."Label-FreeMonitoringof3DTissueModelsviaElectricalImpedanceSpectroscopy."(2019):1-24.
Matysik-Woźniak,Anna,etal."ExaminationofKynurenineToxicityonCornealandConjunctivalEpithelium:InvitroandinvivoStudies."Ophthalmicresearch(2019):1-12.
Compton,Clayton,etal."ReconstitutionoftheVentricularEndocardiumWithinAcellularHearts."RegenerativeEngineeringandTranslationalMedicine(2019):1-11.
Müller,A.L.,etal."4.IdentificationofmiR-301ainPrimaryHumanAtrialFibroblastsandBoneMarrow-DerivedMesenchymalProgenitorCellstoAttenuateEndogenousDifferentiationintoPro-FibroticCells."DifferentiationofPrimaryHumanPro-FibroticMesenchymalCellsInfluencedbyExtracellularMatrixEnvironmentDeterminedbyMicro-RNAExpression(2018):130.
Doblinger,Nina,etal."Impactofhydroxyethylstarchandmodifiedfluidgelatinongranulocytephenotypeandfunction."Transfusion(2019).
Elisabeth,etal."Pro-InflammatoryResponsesinHumanBronchialEpithelialCellsInducedbySporesandHyphalFragmentsofCommonDampIndoorMolds."Internationaljournalofenvironmentalresearchandpublichealth16.6(2019):1085.
Dodmane,PuttappaR.,etal."BiphasicchangesinairwayepithelialcellEGFreceptorbindingandphosphorylationinducedbycomponentsofhogbarndust."Experimentallungresearch44.10(2018):443-454.
McClellan,Alyce,etal."Anovelmechanismfortheprotectionofembryonicstemcellderivedtenocytesfrominflammatorycytokineinterleukin1beta."Scientificreports9(2019).
Wang,Weiling,etal."Aquaporin-3deficiencyslowscystenlargementinexperimentalmousemodelsofautosomaldominantpolycystickidneydisease."TheFASEBJournal(2019):fj-201801338RRR.
Teo,JyeYng,etal."SurfacetetheringofstemcellswithH2O2-responsiveanti-oxidizingcolloidalparticlesforprotectionagainstoxidation-induceddeath."Biomaterials201(2019):1-15.
Gehwolf,Renate,etal."3D-EmbeddedCellCulturestoStudyTendonBIOLOGy."(2019):1-11.
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ProductDisclaimer
ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.
美国AdvancedBioMatrix（简称ABM） www.advancedbiomatrix.comAdvancedBioMatrix（简称ABM）是美国一家著名的生物公司，获得了AllerganInc的授权(Allergan用25年时间不断完善胶原蛋白相关的产品的生产工艺)，将Allergan的专业和技术用于蛋白生产与检测，致力于为组织工程、细胞分析及细胞增殖等研究领域提供优质稳定的产品。AdvancedBioMatrix不断丰富已有产品线，目前可为三维细胞培养提供各种胶原蛋白、纤连蛋白、玻连蛋白、水性凝胶、不同粘度与分子量的透明质酸以及低代成纤维细胞等。在美国全部产品授权Sigma销售。AdvancedBioMatrix是组织培养，细胞分析和细胞增殖三维（3D）应用的生命科学领域的领导者。我们的产品被公认为纯度，功能性和一致性的标准。我们在生产，分离，纯化，冷冻干燥，细胞培养和蛋白质测试，粘附肽，附着因子，底物刚性和其他3D矩阵产品方面拥有丰富的专业知识。我们的专业技术和知识正在被用来确保我们的产品质量最高，批次之间一致且易于为我们的研究客户使用。
美国AdvancedBioMatrix是3D组织培养、细胞检测和细胞增殖等领域实验解决方案的佼佼者。AdvancedBioMatrix在分离、纯化、冻干、细胞培养和蛋白检测、多肽粘附、附着因子、基质硬度和其他3Dmatrix 产品开发方面有着丰富的经验。AdvancedBioMatrix的研发经验和专业知识确保其产品可达到最佳质量，并保证产品之间一致性，方便研究客户使用。以下为AdvancedBioMatrix3DMatrices 产品竞争优势：1. 提供高纯度和成分确定的胞外基质；2. 超过1000余篇文献引用PureCol产品，品质非常均一；3. 在3D培养基领域可提供最全面的产品线；4. 唯一可提供特异性刚性有机硅基板的公司（CytoSoft）;5. 唯一可提供可溶性丝纤蛋白的供应商（可运用于多种3D培养）；6. 如果客户首次接触3D胶原凝胶，AdvancedBioMatrix还是唯一的预制胶原蛋白(PureColEZGel)供应商；
以下产品为AdvancedBioMatrix全球畅销品：1.PureCol 牛源I型胶原蛋白 3mg/ml#5005-100ML2.Nutragen牛源I型胶原蛋白 6mg/ml#5010-50ML3.FibriCol 牛源I型胶原蛋白 10mg/ml#5133-20ML4.VitroCol 人源I型胶原蛋白 #5007-20ML5. 弹性蛋白原 #5052-1MG6.ECMSelectArraykitUltra-36#5170-1EA7.CytoSoft（刚性可变的基底，AdvancedBioMatrix最新添加产品5190-7EA）8. 人III型胶原蛋白 #5021-10MG9. 人IV型胶原蛋白 #5022-5MG10.SilkFibroin溶液 #5154-20ML11.Fibronectin#5080-5MG12.Vitronectin#5051-0.1MG
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