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蚂蚁淘在线 / 品牌中心 / Advanced BioMatrix / 高级生物矩阵/TeloCol®-3//5026-1KIT

高级生物矩阵/TeloCol®-3//5026-1KIT

价格
￥5400.00
货号：5026-1KIT
浏览量：127
品牌：Advanced BioMatrix
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商品描述

ProductDescription

TeloCol®-3bovinecollagensolutionisderivedfromanacidextractionprocessyieldingatelopeptide-intactcollagen.Thepro-peptideregionsatbothendsofthecollagenchain,N-andC-telopeptideregions,aremaintained.

TeloCol®-3collagenisapproximately95%TypeIcollagenwiththeremainderbeingcomprisedofTypeIIIcollagen.Eachproductincludesabottlecontaining50mlofcollagensolutionaccompaniedwithabottleofpre-formulatedneutralizingsolutionfortheformationofacollagengel.Thisproductissuppliedataconcentrationofapproximately3mg/ml.TeloCol®-3issterilefilteredandprovidedinuser-friendlypackagingforuseandstorage.

TeloCol®-3isidealforcoatingofsurfaces,providingpreparationofthinlayersforculturingcells,oruseasasolidgel.

Parameter,Testing,andMethodTeloCol®-3TypeICollagen#5026
SterilizationMethodFiltration
ExtractionMethodAcid-telocollagen
FormSolution
PackageSize50mLKit
StorageTemperatureofCollagen2-10°C
StorageTemperatureofNeutralizationSolutionRoomTemperature
ShelfLifeMinimumof6monthsfromdateofreceipt

CollagenConcentration-Biuret

2.8-3.3mg/mL

CollagenPurity-SilverStaining

>99%
pH1.9-3.0
KineticGelTest(Minutes)<40
GelFormationTubeTest(Minutes)<40

Fibrillogenesis(AbsorbanceUnits)

>0.35

ElectrophoreticPattern-CoomassieBlue

Characteristic
Sterility-USPmodifiedNogrowth
Endotoxin-LAL<10.0EU/mL
Osmolality(mOsmoH2O/kg)<35
CellAttachmentAssayPass
SourceBovineHide
HydrogelYoung"sModulusE(Pa)Characteristic

DirectionsforUse

DownloadthefullPDFversionorcontinuereADIngbelow:

Note:Employasepticpracticestomaintainthesterilityoftheproductthroughoutthepreparationandhandlingofthecollagenandothersolutions.

CoatingProcedure

  1. TransferdesiredvolumeofTeloCol®-3collagensolutionfromthebottletoadilutionvesselifrequired.Furtherdilutetodesiredconcentrationusingsterile0.01NHClsolution.Swirlcontentsgentlyuntilmaterialiscompletelymixed.Atypicalworkingconcentrationmayrangefrom50to100ug/ml.Note:Usetheserecommendationsasguidelinestodeterminetheoptimalcoatingconditionsforyourculturesystem.
  2. AddappropriateamountofdilutedTeloCol®-3collagentotheculturesurfaceensuringthattheentiresurfaceiscoated.
  3. Incubateatroomtemperature,covered,for1-2hours.Aspirateanyremainingmaterial.Alternatively,incubateatroomtemperatureuntilsurfaceisdry.
  4. RinsecoatedsurfacescarefullywithsterilemediumorPBS,avoidscratchingsurfaces.
  5. Coatedsurfacesarereadyforuse.Theymayalsobestoredat2-8°Cdamporairdriedifsterilityismaintained.

3-DGelPreparationProcedureusingthesuppliedNeutralizationSolution

Note:Itisrecommendedthatthecollagenandotherworkingsolutionsbechilledandkeptoniceduringthepreparationofthecollagen.

  1. Determinethedesiredvolumeofcollagenrequired.
  2. Transfer1partofchilledneutralizationsolutionintoasterilemixingvesselortube.
  3. Transfer9partsoftheTeloCol®-3collagenintothesterilemixingvesselortubeforatotalof10parts.
  4. Gentlyagitatethemixtureorpipetupanddowntomix.Vortexingisnotrecommended.
  5. Dispensethecollagenmixtureinthedesiredsterileplatesorculturevessels.
  6. Incubateat37°Cfor1hourforgelformation.

3-DGelPreparationProcedurewithoutusingthesuppliedNeutralizationSolution

Note:Itisrecommendedthatthecollagenandotherworkingsolutionsbechilledandkeptoniceduringthepreparationofthecollagen.

  1. Slowlyadd1partofchilled10XPBSor10Xculturemediato8partsofchilledcollagensolutionwithgentleswirling.
  2. AdjustpHofmixtureto7.2–7.6usingsterile0.1MNaOH.MonitorpHadjustmentcarefully(pHmeter,phenolred,orpHpaper).Mixwithgentleswirling.
  3. Adjustfinalvolumetoatotalof10partswithsterilewaterandmix.
  4. Topreventgelation,maintaintemperatureofmixtureat2–10°C.
  5. DispensetheTeloCol®-3collagenmixtureinthedesiredsterileplatesorculturevessels.
  6. Toformgel,warmto37°C.Allowapproximately30to60minutesforgelformation.

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

ReferencesforTeloCol®:

Drzewiecki,KathrynE.,etal.“CircularDichroismSpectroscopyofCollagenFibrillogenesis:ANewUseforanOldTechnique.”BiophysicalJournal,vol.111,no.11,2016,pp.2377–2386.,doi:10.1016/j.bpj.2016.10.023

Wingender,Brian,etal.“BiomimeticOrganizationofCollagenMatricestoTemplateBone-likeMicrostructures.”MatrixBIOLOGy,vol.52-54,2016,pp.384–396.,doi:10.1016/j.matbio.2016.02.004.

Burla,F.,Tauber,J.,Dussi,S.,Gucht,J.V.D.&Koenderink,G.H.Stressmanagementincompositebiopolymernetworks.NaturePhysics15,549–553(2019).

ProductCertificateofAnalysis

Noresultfor.

ProductVideos

link to library blog - Telo vs Atelo Collagen
TelovsAteloCollagen

Video

link to library blog - Neutralization Solution for Easy 3D Collagen Gels
NeutralizationSolutionforEasy3DCollagenGels

Video

link to library blog - Collagen Concentration vs Gel Stiffness
CollagenConcentrationvsGelStiffness

Video

link to library blog - 4 Key Components for 3D Collagen Gels
4KeyComponentsfor3DCollagenGels

Video

link to library blog - How to Make 3D Collagen Hydrogels
HowtoMake3DCollagenHydrogels

Video

link to library blog - Seeding Collagen Gels with Cells
SeedingCollagenGelswithCells

Video

link to library blog - Coating a Glass Coverslip with Collagen
CoatingaGlassCoverslipwithCollagen

Video

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SafetyandDocumentation

SafetyDataSheet

CertificateofOrigin

DeclarationofMaterialSource

ProductDisclaimer

ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.

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