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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/VitroCol®//5007-20ML
产品编号:5007-20ML
市  场 价:¥6200.00
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:$310.00
品      牌: Advanced BioMatrix
公      司:Advanced BioMatrix ,Inc.
公司分类:
Advanced BioMatrix/VitroCol®//5007-20ML
商品介绍

ProductDescription

VitroCol®collagenisthefirstwidelyavailable,naturallyproducedpurifiedhumancollagenforresearchpurposes.VitroCol®setsthestandardforpurity(>99%collagencontent),functionalityandrepresentstheonlynative-likehumancollagenoffered.

VitroCol®collagenisnaturallysecretedfromhumanneo-natalfibroblastcells.Thehumanfibroblastsareculturedinoptimalconditionsallowingthefibroblasttonaturallyandefficientlysecretextracellularmatrix.Theextracellularmatrixisthenprocessedandpurifiedtoyieldthenaturallyproducedhumancollagen.

VitroCol®isapproximately97%TypeIhumancollagenwiththeremainderbeingcomprisedofTypeIIIcollagen.VitroCol®issuppliedatapproximately3mg/mlconcentration.TheconcentrationforeachspecificlotisprovidedonaCertificateofAnalysisthatisavailablewiththepurchaseofeachproduct.VitroCol®issolubleatelocollagenin0.01NHCI,therefore,thepHis2.

VitroCol®isespeciallyidealforhumancellculturesystemswhencoatingofsurfaces,providingpreparationsofthinlayersofculturingcells,oruseasasolidgel.VitroCol®humancollagenisprovidedinuser-friendlypackagingforuseandstorage.VitroCol®issterilefilteredandissuppliedasareadytousesolution.

Parameter,Testing,andMethodVitroCol®TypeICollagen#5007
SterilizationMethodFiltration
ExtractionMethodEnzyme-atelocollagen
FormSolution
PackageSize20mL,100mL
StorageTemperature2-10°C
ShelfLifeMinimumof6monthsfromdateofreceipt

CollagenConcentration-Biuret

2.9-3.2mg/mL

CollagenPurity-SilverStaining

>99%
pH1.9-2.1
KineticGelTest(Minutes)<40
GelFormationTubeTest(Minutes)<40

Fibrillogenesis(AbsorbanceUnits)

>0.5

ElectrophoreticPattern-CoomassieBlue

Characteristic
Sterility-USPmodifiedNogrowth
Endotoxin-LAL<5.0EU/mL
Osmolality(mOsmoH2O/kg)<35
CellAttachmentAssayPass
SourceHumanNeo-NatalFibroblasts
HydrogelYoung"sModulusE(Pa)Characteristic

DirectionsforUse

DownloadthefullPDFversionorcontinuereADIngbelow:

CoatingProcedure

Note:Usetheserecommendationsasguidelinestodeterminetheoptimalcoatingconditionsforyourculturesystem.

  1. Transferdesiredvolumeofcollagensolutionfromthebottletoadilutionvesselasrequired.Dilutetodesiredconcentrationusingsterile0.01NHClsolution.Atypicalworkingconcentrationmayrangefrom50to100ug/ml.
  2. AddappropriateamountofdilutedVitroCol®materialtotheculturesurface.
  3. Incubateatroomtemperature,covered,for1-2hours.Aspirateanyremainingmaterial.Alternatively,incubateatroomtemperatureuntilsurfaceisdry.
  4. RinsecoatedsurfacescarefullywithsterilemediumorPBS,avoidscratchingsurfaces.
  5. Coatedsurfacesarereadyforuse.Theymayalsobestoredat2-8°Cdamporairdriedifsterilityismaintained.

3-DGelPreparationProcedure

  1. Slowlyadd1partofchilled10XPBSor10Xculturemediato8partsofchilledcollagensolutionwithgentleswirling.
  2. AdjustpHofmixtureto7.0-7.5usingsterile0.1MNaOH.MonitorpHadjustmentcarefully(pHmeter,phenolred,orpHpaper)
  3. Adjustfinalvolumetoatotalof10partswithsterilewater.
  4. Topreventgelation,maintaintemperatureofmixtureat2–10°C.
  5. Toformgel,warmto37°C.Allowapproximately90to120minutesforgelformation.

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

ReferencesforVitroCol®:

Andrée,B.etal.Formationofthree-dimensionaltubularendothelialcellnetworksunderdefinedserum-freecellcultureconditionsinhumancollagenhydrogels.ScientificReports9,(2019).

Shieh,HesterF.,etal."ComparisonsofhumanamnioticmesenchymalstemcellviABIlityinFDA-approvedcollagen-basedscaffolds:Implicationsforengineereddiaphragmaticreplacement."Journalofpediatricsurgery52.6(2017):1010-1013.

vanderVelden,JosLJ,etal."TransformingGrowthFactor-ßInducesAMesenchymalProfileInHumanNasalEpithelialCells."D76.ALVEOLAREPITHELIUM:NOVELTOOLSANDPHENOTYPES.AmericanThoracicSociety,2012.A6322-A6322.

Tashima,Takumi,etal."Osteomodulinregulatesdiameterandaltersshapeofcollagenfibrils."Biochemicalandbiophysicalresearchcommunications463.3(2015):292-296.

Spiller,KaraL.,SuzanneA.Maher,andAnthonyM.Lowman."Hydrogelsfortherepairofarticularcartilagedefects."TissueengineeringpartB:reviews17.4(2011):281-299.

Brilha,Sara,etal."Monocyteadhesion,migration,andextracellularmatrixbreakdownareregulatedbyintegrinαVβ3inMycobacteriumtuberculosisinfection."TheJournalofImmunology199.3(2017):982-991.

Jonsdottir,HuldaR.,andRonaldDijkman."Characterizationofhumancoronavirusesonwell-differentiatedhumanairwayepithelialcellcultures."Coronaviruses.HumanaPress,NewYork,NY,2015.73-87.

Sabbione,Florencia,etal."Neutrophilextracellulartrapsstimulateproinflammatoryresponsesinhumanairwayepithelialcells."Journalofinnateimmunity9.4(2017):387-402.

DosSantosBrilha,S.,etal."Monocyteadhesion,migrationandextracellularmatrixbreakdownisregulatedbyintegrinαVβ3inMycobacteriumtuberculosisinfection."

Brilha,Sara,etal."MonocyteAdhesion,Migration,andExtracellularMatrixBreakdownIsRegulatedbyIntegrinaVb3inMycobacteriumtuberculosisInfection."(2017).

Colace,T.,etal."Analysisofmorphologyofplateletaggregatesformedoncollagenunderlaminarbloodflow."Annalsofbiomedicalengineering39.2(2011):922-929.

Muthard,RyanW.,andScottL.Diamond."Bloodclotsarerapidlyassembledhemodynamicsensors:flowarresttriggersintraluminalthrombuscontraction."Arteriosclerosis,thrombosis,andvascularBIOLOGy32.12(2012):2938-2945.

Maloney,S.F.,LawrenceF.Brass,andS.L.Diamond."P2Y12orP2Y1inhibitorsreduceplateletdepositioninamicrofluidicmodelofthrombosiswhileapyraselacksefficacyunderflowconditions."IntegrativeBiology2.4(2010):183-192.

Bauer,RebeccaN.,etal."Interactionwithepithelialcellsmodifiesairwaymacrophageresponsetoozone."Americanjournalofrespiratorycellandmolecularbiology52.3(2015):285-294.

Kawamura,Shunsuke,etal."IdentificationofcommonmonocyteProgenitors,pre-monocytes,andgranulocytemonocyteprogenitorsinhumanumbilicalcordblood."ExperimentalHematology43.9(2015):S72.

ProductCertificateofAnalysis

Noresultfor.

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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