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胶原蛋白、纤连蛋白、玻连蛋白、水性凝胶、低代成纤维细胞
℡ 4000-520-616
℡ 4000-520-616
Advanced BioMatrix/RatCol® for 3D Hydrogels//5153-1KIT
产品编号:5153-1KIT
市  场 价:¥3200.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$160.00
品      牌: Advanced BioMatrix
公      司:Advanced BioMatrix ,Inc.
公司分类:
Advanced BioMatrix/RatCol® for 3D Hydrogels//5153-1KIT
商品介绍

ProductDescription

RatCol®TypeIAcidSolubleRatTailCollagencontains100mgataconcentrationofapproximately4mg/mLina0.02Maceticacidsolution(pH2to3).RatCol®collagenissolubletelo-collagen.Eachproductincludesabottlecontaining100mgofcollagensolutionaccompaniedwithabottleofpre-formulatedneutralizingsolutionfortheformationofacollagengel.Thiscollagenproductisprovidedinuser-friendlypackagingforuseandstorage.Thisproductissterilefilteredandissuppliedasareadytousesolution.

Thisproductisidealforcoatingofsurfaces,providingpreparationofthinlayersforculturingcells,oruseasasolidgel.RatCol®collagenissuitableforapplicationsusingavarietyofcelllinesincludinghepatocytes,fibroblastsandepithelialcells.

Parameter,Testing,andMethodRatCol®TypeICollagen#5153
SterilizationMethodFiltration
ExtractionMethodAcid-telocollagen
FormSolution
PackageSize100mg(~25mL)Kit
StorageTemperatureofCollagen2-10°C
StorageTemperatureofNeutralizationSolutionRoomTemperature
ShelfLifeMinimumof6monthsfromdateofreceipt

CollagenConcentration-Biuret

3.5-4.5mg/mL

CollagenPurity-SilverStaining

>99%
pH3.0-3.8
KineticGelTest(Minutes)<40

GelFormationTubeTest(Minutes)

<40

ElectrophoreticPattern-CoomassieBlue

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

DirectionsforUse

DownloadthefullPDFversionorcontinuereADIngbelow:

CoatingProcedure

Note:Employasepticpracticestomaintainthesterilityoftheproductthroughoutthepreparationandhandlingofthecollagenandothersolutions.

  1. Transferdesiredvolumeofcollagensolutionfromthebottletoadilutionvesselifrequired.Furtherdilutetodesiredconcentrationusingsterile0.1%aceticacidsolution.Atypicalworkingconcentrationmayrangefrom50to100ug/mL.Note:Usetheserecommendationsasguidelinestodeterminetheoptimalcoatingconditionsforyourculturesystem.
  2. AddappropriateamountofdilutedRatTailcollagentotheculturesurface.
  3. Incubateatroomtemperature,covered,for1-2hours.Aspirateanyremainingmaterial.Alternatively,incubateatroomtemperatureuntilsurfaceisdry.
  4. RinsecoatedsurfacescarefullywithsterilemediumorPBS,avoidscratchingsurfaces.
  5. Coatedsurfacesarereadyforuse.Theymayalsobestoredat2-8°Cdamporairdriedifsterilityismaintained.

3-DGelPreparationProcedureUsingtheSuppliedNeutralizationSolution

Note:Employasepticpracticestomaintainthesterilityoftheproductthroughoutthepreparationandhandlingofthecollagenandothersolutions.

Note:Itisrecommendedthatthecollagenandotherworkingsolutionsbechilledandkeptoniceduringthepreparationofthecollagen.

  1. Determinethedesiredvolumeofcollagenrequired.
  2. Transfer1partofchilledneutralizationsolutionintoasterilemixingvesselortube.
  3. Transfer9partsoftheRatTailCollagenintothesterilemixingvesselortubeforatotalof10parts.
  4. Gentlyagitatethemixtureorpipetupanddowntomix.Vortexingisnotrecommended.
  5. DispensetheRatTailcollagenmixtureinthedesiredsterileplatesorculturevessels.
  6. Incubateat37°Cfor1hourforgelformation.

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

ReferencesforRatCol®:

Dollinger,BryanR.,etal."Reactiveoxygenspeciesshieldinghydrogelforthedeliveryofadherentandnonadherenttherapeuticcelltypes."TissueEngineeringPartA23.19-20(2017):1120-1131.

Jia,Hong,etal."Thetumorcell‐secretedmatricellularproteinWISP1drivespro‐metastaticcollagenlinearization."TheEMBOJournal(2019).

PérezCardona,DavidJosé."Phenotypeanalysisofdermal-epidermalorganotypicsmadewithhacatcelllineseedingontopofthreehumanfibroblastpopulatedmatrices(polyethyleneterephthalate,fibrinorcollagenI+III)."(2017).

Karki,SuryaB.,etal."Investigationofnon-thermalplasmaeffectsonlungcancercellswithin3Dcollagenmatrices."JournalofPhysicsD:AppliedPhysics50.31(2017):315401.

Keating,M.,etal."Spatialdistributionsofpericellularstiffnessinnaturalextracellularmatricesaredependentoncell-mediatedproteolysisandcontractility."ActaBiomaterialia57(2017):304-312.

Blum,KevinM.,etal."Acellularandcellularhigh-density,collagen-fibrilconstructswithsuprafibrillarorganization."Biomaterialsscience4.4(2016):711-723.

Kaufman,Gili,andDragoSkrtic."Spatialdevelopmentofgingivalfibroblastsanddentalpulpcells:Effectofextracellularmatrix."TissueandCell49.3(2017):401-409.

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

SeeMore

SafetyandDocumentation

SafetyDataSheet

CertificateofOrigin

ProductDisclaimer

ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.

品牌介绍
美国Advanced BioMatrix(简称ABM) www.advancedbiomatrix.com
Advanced BioMatrix(简称ABM)是美国一家著名的生物公司,获得了Allergan Inc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。

Advanced BioMatrix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。

Advanced BioMatrix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。

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