ProductDescriptionPureCol®EZGelisaready-to-usecollagensolutionthatformsafirmgelbysimplywarmingto37°Cinanincubator.TheproductconsistsofpurifiedTypeIcollagenataconcentrationof0.5%,(~5mg/ml),DMEM/F-12mediumandamixtureofL-glutamineanddipeptide(L-alanine-L-glutamine)toprovidealong-lastingL-glutaminesourceforcellculture.PureCol®EZGelisdesignedtoimprovegelconsistencybypre-formulationofthemediumandadjustmentoftheproducttoaneutralpH.Thisproductavoidstheinconsistenciesinthepreparationofthegelthatcanarisethroughvariablesofreagentaddition,pHadjustmentandhandlingconditions.PureCol®EZGelisidealforprovidingafirmgelandcanalsobeusedinthepreparationofathinlayerforculturingcells.PureCol®EZGelcollagenisprovidedinauser-friendlypackagingforuseandstorage.Thisproductissterileandsuppliedasareadytousesolution.Parameter,Testing,andMethodPureCol®EZGelTypeICollagen#5074SterilizationMethodFiltrationExtractionMethodEnzyme-atelocollagenFormSolutionPackageSize35mLStorageTemperature2-10°CShelfLifeMinimumof6monthsfromdateofreceiptCollagenConcentration-Biuret~5mg/mLCollagenPurity-SilverStaining>99%pH6.9-7.4KineticGelTest(Minutes)<40GelFormationTubeTest(Minutes)<40Fibrillogenesis(AbsorbanceUnits)>0.5ElectrophoreticPattern-CoomassieBlueCharacteristicSterility-USPmodifiedNogrowthEndotoxin-LAL<1.0EU/mLOsmolality(mOsmoH2O/kg)300-360CellAttachmentAssayPassSourceBovineHideHydrogelYoung"sModulusE(Pa)CharacteristicMediumSupplementDMEM/F-12L-GlutamineSourceMixtureofL-GlutamineandDipeptide(L-alanine-L-glutamine)DirectionsforUseDownloadthefullPDFversionorcontinuereADIngbelow:3-DPreparationProcedureRemovePureCol®EZGelfrom2–10°Cstorage.Important:Topreventgelation,maintaintemperatureofproductat2–10°C.IntroducePureCol®EZGelintocellculturesystem.CellscanbeaddedtothePureCol®EZGelsolution.Toformgel,warmto37°C.Thebeginningofgelationwilloccurwithin40minutes,butallowapproximately90tominutesforfirmgelformation.ProductQ&AWhatistheconcentrationofPureCol®EZGel?Theconcentrationrangesfrom5.2-5.5mg/ml.IsthereDNAinthecollagenproducts?WecompletedastudytoshowthatDNAiscompletelydestroyedatpH2,anddemonstratedthatourcollagenproductsdonotcontainDNA.HowistheAminoAcidAnalysisperformed?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.Canyoudilutecollagenwith1XPBSfor2Dcoatings?Dilutingwith1XPBS(ratherthanwateror0.01NHCl)wouldhaveaneffectforcoatingpurposes.ItwouldchangethepHofthedilutedcollagensolutionfromacidtoneutralpH.ThepHchangewilltransformthecollagenmoleculesfromamolecularformtoafibrillarform;andthenthenatureofcoatingsurfacewillbechangedfromamonomericcoatingtoafibrillarcoating.WhatantibodiesdoyourecommendforyourTypeIcollagenproducts?WeusethefollowingantibodiesfromSouthernBiotech:1.1310-02–GoatAnti-TypeICollagen-FITC2.1310-08–GoatAnti-TypeICollagen-BIOT3.7100-05–Streptavidin-HRPWhatisthethicknessofthe‘molecularcollagen’attachingtothepolystyrenesurfaceofcultureware?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.Whatisthechargeofyourcollagenproducts?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.Whatisthemonomertooligomerratioinyourcollagenproducts?Usingrotaryshadowingtechniqueundertransmissionelectronmicroscopy,itwasfoundthatourcollagen,onaverage,consistsofapproximately80%monomers,13%dimers,trimers,andoligomerswiththeremaining7%collagenfragments.IspepsintreatedcollagenstillMMP-Sensitive?Yes.ThecollagenmoleculeinPureCol,Nutragen,VitroCol,andallofourotherAtelocollagenproductswerepreparedfromnativecollagenmatrixbypepsintreatmentundercontrolledconditionstoremovethenon-helicalportion,telo-peptides,onlyandthehelicalportionisintact.Inthiscase,theenzymaticactivesitesforMMP(MatrixMetalloproteinase),suchasforMammalianCollagenaseMatrixMetalloproteinase8(MMP-8),onthemoleculewaspreserved.Thesepepsintreatedcollagenproductsshouldbehaveasnativeintactcollagen.IsthereTGFBetainthecollagenproducts?TGFbetawouldhavebeendigestedwiththepepsinenzymaticdigestionstep.ItwasundetectablebySDSPAGEsilverstainaswell.Wedidn’tdoanyspecificmeasurementsbyELISAhoweverbutpresencesofTGFbetaisnotanticipated.Howdoyoudetermineproteinconcentration?WeprimarilyusetheBiuretmethod,butwealsouseBCA,AAA,andhydroxyl-prolineassays.IsmycollagenOKifitwasfrozen,orleftoutatroomtemperature?-Collagensolutionsthatarefrozentendtohaveissuesforming3Dhydrogels,andwilllikelynotwork.Thesolutionsshouldstillbegoodfor2Dcoatings.-Collagensolutionsthatareleftoutatroomtemperatureforextendedperiodsoftimemayshowsignsofdegradation,whichwillaffecttheformationof3Dhydrogels.Itislikelystillfinefor2Dcoatings.Ourrecommendationisthis:Ifyouareusingtheproductdirectlyforapublication,wehighlysuggestbuyinganewbottleiftheoneyouhavewascompromised.ProductReferencesReferencesforPureCol®EZGel:Senior,J.J."FabricationofComplexHydrogelStructuresUsingSuspendedLayerAdditiveManufacturing(SLAM)."AdvancedFunctionalMaterials1904845(2019).doi:10.1002/adfm.201904845Yang,Guang,etal."Enhancementoftenogenicdifferentiationofhumanadiposestemcellsbytendon-derivedextracellularmatrix."Biomaterials34.37(2013):9295-9306.Calvao,DominickJoseph,GaetanaA.D"Alesio-Spina,andPatrickEdwardThomas."FabricationofaNutrientPerfusionEnhancingCartilageTissueScaffold."(2015).TracyLindquist,DougJones,JohnJackman2,ShannonHostetter,andJesseHostetter.EvaluatingtheinvivoimmuneresponsetoMycobacteriumaviumsubspeciesparatuberculosisinfectioninnaïveandvaccinatedcalves1001(2018):26.Moxon,SamuelR.,etal."Blendedalginate/collagenhydrogelspromoteneurogenesisandneuronalmaturation."MaterialsScienceandEngineering:C(2019):109904.Gehwolf,Renate,etal."3D-EmbeddedCellCulturestoStudyTendonBIOLOGy."(2019):1-11.Padilla‐Martinez,JuanPablo,RuishengWang,andWalfreFranco."Evaluationofcellandmatrixmechanicsusingfluorescenceexcitationspectroscopy:Feasibilitystudyincollagengelscontainingfibroblasts."Lasersinsurgeryandmedicine48.4(2016):377-384.Zhou,C.,etal."BMP4promotesvascularizationofhumanadiposestromalcellsandendothelialcellsinvitroandinvivo."Cellproliferation46.6(2013):695-704.Gehwolf,Renate,etal."3D-EmbeddedCellCulturestoStudyTendonBiology."(2019):1-11.Foramorecomprehensivelistofreferences,clickhere.ProductCertificateofAnalysisNoresultfor.ProductVideosPureCol®EZGel-Easy3DCollagenGelsVideoSeeMore>TelovsAteloCollagenVideoSeeMore>SeedingCollagenGelswithCellsVideoSeeMore>4KeyComponentsfor3DCollagenGelsVideoSeeMore>CollagenConcentrationvsGelStiffnessVideoSeeMore>SeeMoreSafetyandDocumentationSafetyDataSheetCertificateofOriginDeclarationofMaterialSourceProductDisclaimerThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.

美国Advanced BioMatrix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。Advanced BioMatrix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3D matrix 产品开发方面有着丰富的经验。Advanced BioMatrix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Advanced BioMatrix 3D Matrices 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，Advanced BioMatrix还是唯一的预制胶原蛋白(PureCol EZ Gel)供应商。

以下产品为Advanced BioMatrix全球畅销品：
1. PureCol 牛源I型胶原蛋白 3 mg/ml #5005-100ML
2. Nutragen牛源I型胶原蛋白 6 mg/ml #5005-100ML
3. FibriCol 牛源I型胶原蛋白 10 mg/ml #5133-20ML
4. VitroCol 人源I型胶原蛋白 #5007-20ML
5. 弹性蛋白原 #5052-1MG
6. ECM Select Array kit Ultra-36 #5170-1EA
7. CytoSoft（刚性可变的基底，Advanced BioMatrix最新添加产品5190-7EA）
8. 人III型胶原蛋白 #5021-10MG
9. 人IV型胶原蛋白 #5022-5MG
10. Silk Fibroin溶液 #5154-20ML
11. Fibronectin #5080-5MG
12. Vitronectin #5051-0.1MG

Advanced BioMatrix最畅销的产品是PureCol（5005-100ML）。目前已有上千发表的论文使用这款产品。（Nutragen 5010、fibricol 5133和PureCol的物质成分一样，仅仅是浓度不同）浓度越高，胶原蛋白硬度越强。