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Advanced BioMatrix/FibriCol®//5133-20ML

ProductDescription

FibriCol®collagenisknownasthestandardofallcollagensforpurity(>99%collagencontent),functionality,andthemostnative-likecollagenavailable.FibriCol®isisolatedfrombovinehidessourcedfromtheonlycontrolled,closedherdintheUnitedStates.AdvancedBiomatrix’smanufacturingprocessescomplywithstringentqualitystandardsthathaveproventoyieldunsurpassedlot-to-lotconsistency.

FibriCol®collagenisapproximately97%TypeIcollagenwiththeremainderbeingcomprisedofTypeIIIcollagen.FibriCol®collagenissuppliedatapproximatelya10mg/mLconcentration.TheconcentrationforeachspecificlotisprovidedonaCertificateofAnalysisthatisavailablewiththepurchaseofeachproduct.FibriCol®issolubleatelocollagenin0.01NHCI,therefore,thepHis2.

FibriCol®collagenisidealforapplicationswherehighconcentrationsofcollagenareneeded.FibriCol®collagenisprovidedina20mLvolumeandiscontainedinuser-friendlypackagingforuseandstorage.FibriCol®issterilefilteredandissuppliedasareadytousesolution.

Parameter,Testing,andMethodFibriCol®TypeICollagen#5133
SterilizationMethodFiltration
ExtractionMethodEnzyme-atelocollagen
FormSolution
PackageSize20mL,100mL
StorageTemperature2-10°C
ShelfLifeMinimumof6monthsfromdateofreceipt

CollagenConcentration-Biuret

9.0-11.5mg/mL

CollagenPurity-SilverStaining

>99.9%
pH1.9-2.1
KineticGelTest(Minutes)<40
GelFormationTubeTest(Minutes)<40

Fibrillogenesis(AbsorbanceUnits)

>0.5

ElectrophoreticPattern-CoomassieBlue

Characteristic
Sterility-USPmodifiedNogrowth
Endotoxin-LAL<1.0EU/mL
Osmolality(mOsmoH2O/kg)<35
CellAttachmentAssayPass
SourceBovineHide
HydrogelYoung"sModulusE(Pa)Characteristic

DirectionsforUse

DownloadthefullPDFversionorcontinuereADIngbelow:

3-DGelPreparationProcedure

  1. Slowlyadd1partofchilled10XPBSor10Xculturemediato8partsofchilledcollagensolutionwithgentlemixing/swirling.
  2. AdjustpHofmixtureto7.2–7.6.Useofsterile0.1MNaOHisrecommended.MonitorpHadjustmentcarefully(pHmeter,phenolred,orpHpaper).
  3. Adjustfinalvolumetoatotalof10partswithsterilewater.
  4. Topreventgelation,maintaintemperatureofmixtureat2-10°C.
  5. Toformgel,warmto37°C.Forbestresultsallowapproximately90minutesforgelformation

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

ReferencesforFibriCol®:

Berger,AnthonyJ.,etal."Decouplingtheeffectsofstiffnessandfiberdensityoncellularbehaviorsviaaninterpenetratingnetworkofgelatin-methacrylateandcollagen."Biomaterials141(2017):125-135.

Kreimendahl,Franziska,etal."Three-dimensionalprintingandangiogenesis:tailoredagarose-typeIcollagenblendscomprisethree-dimensionalprintABIlityandangiogenesispotentialfortissue-engineeredsubstitutes."TissueEngineeringPartC:Methods23.10(2017):604-615.

Mas-Vinyals,Anna,etal."Improvinglinkinginterfacebetweencollagen-basedhydrogelsandbone-likesubstrates."ColloidsandSurfacesB:Biointerfaces(2019).

Schoppmeyer,Rouven,etal."Light-sheetmicroscopyforthree-dimensionalvisualizationofhumanimmunecells."JoVE(JournalofVisualizedExperiments)136(2018):e57651.

Semina,E.V.,etal."Three-dimensionalmodelofbiomatrixasamethodofstudyingbloodvesselsandnervegrowthintissueengineeringstructures."MoscowUniversityChemistryBulletin71.3(2016):172-177.

Pompilio,Arianna,etal."MyroidesodoratimimusFormsStructurallyComplexandInherentlyAntibiotic-ResistantBiofilminaWound-LikeinvitroModel."FrontiersinmicroBIOLOGy8(2017):2591.

Christian,SherriL.,etal."Collagenoverlayscaninhibitleptinandadiponectinsecretionbutnotlipidaccumulationinadipocytes."PeerJ6(2018):e4641.

Vining,KyleH.,AlexanderStafford,andDavidJ.Mooney."Sequentialmodesofcrosslinkingtuneviscoelasticityofcell-instructivehydrogels."Biomaterials188(2019):187-197.

Kreimendahl,F.,etal."Combinationofvascularizationandciliaformationforthree‐dimensionalairwaytissueengineering."JournalofBiomedicalMaterialsResearchPartA(2019).

Chen,Yin-Quan,etal."Earlystagemechanicalremodelingofcollagensurroundingheadandnecksquamouscellcarcinomaspheroidscorrelatesstronglywiththeirinvasioncapability."Actabiomaterialia84(2019):280-292.

Caliari,StevenR.,andJasonA.Burdick."Apracticalguidetohydrogelsforcellculture."Naturemethods13.5(2016):405.

Szulc,DanielAndrzej,andHai‐LingMargaretCheng."One‐StepLabelingofCollagenHydrogelswithPolydopamineandManganesePorphyrinforNon‐InvasiveScaffoldTrackingonMagneticResonanceImaging."Macromolecularbioscience(2019):1800330.

ProductCertificateofAnalysis

Noresultfor.

ProductVideos

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SafetyandDocumentation

SafetyDataSheet

CertificateofOrigin

DeclarationofMaterialSource

ProductDisclaimer

ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.

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