ProductDescription
PureCol®CollagenCoated,100X20mmDisheshaveauniformandconsistentapplicationofhighqualityTypeIcollagenonclearpolystyrenesurface.Plateshavearaisedringaroundthecircumference(SureGrip)ofthedishthatprovidesasecuregripofboththedishandlid.Embossedarrowsonthedishandlidprovideorientationforcellsiteobservation.Dishesareasepticallyprocessedandarenon-pyrogenic.Dishesareavailableinquantitiesof10dishespersleeveor50dishespercase.
| Parameter,Testing,andMethod | PureCol®Coated100mmDishes#5028 |
| SterilizationMethod | AsepticallyProcessed |
| Size | 100x20mmDish |
| CollagenUsed | PureCol®TypeI |
| Stackable | Yes |
| VentilationCams | Yes |
| StorageTemperature | 2-30°C |
QuantityperPackage | 10Dishes |
| ShelfLife | Minimumof6monthsfromdateofreceipt |
| FlaskPolymer | Polystyrene |
| TisueCultureTreatedPriortoCoating | Yes |
| GrowthAreaperDish | 58cm2 |
| TypicalWorkingVolumeperDish | 10to20mL |
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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ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.
