ProductDescription
| Parameter,Testing,andMethod | PureCol®EZGelTypeICollagen#5074 |
| SterilizationMethod | Filtration |
| ExtractionMethod | Enzyme-atelocollagen |
| Form | Solution |
| PackageSize | 35mL |
| StorageTemperature | 2-10°C |
| ShelfLife | Minimumof6monthsfromdateofreceipt |
CollagenConcentration-Biuret | ~5mg/mL |
CollagenPurity-SilverStaining | >99% |
| pH | 6.9-7.4 |
| KineticGelTest(Minutes) | <40 |
| GelFormationTubeTest(Minutes) | <40 |
Fibrillogenesis(AbsorbanceUnits) | >0.5 |
ElectrophoreticPattern-CoomassieBlue | Characteristic |
| Sterility-USPmodified | Nogrowth |
| Endotoxin-LAL | <1.0EU/mL |
| Osmolality(mOsmoH2O/kg) | 300-360 |
| CellAttachmentAssay | Pass |
| Source | BovineHide |
| HydrogelYoung"sModulusE(Pa) | Characteristic |
| MediumSupplement | DMEM/F-12 |
| L-GlutamineSource | MixtureofL-GlutamineandDipeptide(L-alanine-L-glutamine) |
DirectionsforUse
DownloadthefullPDFversionorcontinuereADIngbelow:
3-DPreparationProcedure
- RemovePureCol®EZGelfrom2–10°Cstorage.Important:Topreventgelation,maintaintemperatureofproductat2–10°C.
- IntroducePureCol®EZGelintocellculturesystem.CellscanbeaddedtothePureCol®EZGelsolution.
- Toformgel,warmto37°C.Thebeginningofgelationwilloccurwithin40minutes,butallowapproximately90tominutesforfirmgelformation.
ProductQ&A
Theconcentrationrangesfrom5.2-5.5mg/ml.
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
ReferencesforPureCol®EZGel:
Senior,J.J."FabricationofComplexHydrogelStructuresUsingSuspendedLayerAdditiveManufacturing(SLAM)."AdvancedFunctionalMaterials1904845(2019).doi:10.1002/adfm.201904845
Yang,Guang,etal."Enhancementoftenogenicdifferentiationofhumanadiposestemcellsbytendon-derivedextracellularmatrix."Biomaterials34.37(2013):9295-9306.
Calvao,DominickJoseph,GaetanaA.D"Alesio-Spina,andPatrickEdwardThomas."FabricationofaNutrientPerfusionEnhancingCartilageTissueScaffold."(2015).
TracyLindquist,DougJones,JohnJackman2,ShannonHostetter,andJesseHostetter.EvaluatingtheinvivoimmuneresponsetoMycobacteriumaviumsubspeciesparatuberculosisinfectioninnaïveandvaccinatedcalves1001(2018):26.
Moxon,SamuelR.,etal."Blendedalginate/collagenhydrogelspromoteneurogenesisandneuronalmaturation."MaterialsScienceandEngineering:C(2019):109904.
Gehwolf,Renate,etal."3D-EmbeddedCellCulturestoStudyTendonBIOLOGy."(2019):1-11.
Padilla‐Martinez,JuanPablo,RuishengWang,andWalfreFranco."Evaluationofcellandmatrixmechanicsusingfluorescenceexcitationspectroscopy:Feasibilitystudyincollagengelscontainingfibroblasts."Lasersinsurgeryandmedicine48.4(2016):377-384.
Zhou,C.,etal."BMP4promotesvascularizationofhumanadiposestromalcellsandendothelialcellsinvitroandinvivo."Cellproliferation46.6(2013):695-704.
Gehwolf,Renate,etal."3D-EmbeddedCellCulturestoStudyTendonBiology."(2019):1-11.
Foramorecomprehensivelistofreferences,clickhere.
ProductCertificateofAnalysis
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SafetyandDocumentation
SafetyDataSheet
CertificateofOrigin
DeclarationofMaterialSource
ProductDisclaimer
ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.
美国AdvancedBioMatrix（简称ABM） www.advancedbiomatrix.comAdvancedBioMatrix（简称ABM）是美国一家著名的生物公司，获得了AllerganInc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。AdvancedBioMatrix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。AdvancedBioMatrix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。
美国AdvancedBioMatrix是3D组织培养、细胞检测和细胞增殖等领域实验解决方案的佼佼者。AdvancedBioMatrix在分离、纯化、冻干、细胞培养和蛋白检测、多肽粘附、附着因子、基质硬度和其他3Dmatrix 产品开发方面有着丰富的经验。AdvancedBioMatrix的研发经验和专业知识确保其产品可达到最佳质量，并保证产品之间一致性，方便研究客户使用。以下为AdvancedBioMatrix3DMatrices 产品竞争优势：1. 提供高纯度和成分确定的胞外基质；2. 超过1000余篇文献引用PureCol产品，品质非常均一；3. 在3D培养基领域可提供最全面的产品线；4. 唯一可提供特异性刚性有机硅基板的公司（CytoSoft）;5. 唯一可提供可溶性丝纤蛋白的供应商（可运用于多种3D培养）；6. 如果客户首次接触3D胶原凝胶，AdvancedBioMatrix还是唯一的预制胶原蛋白(PureColEZGel)供应商；
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浓度的测定，需测定在230、260和280nm处的消光值，经验数据表明，纯净的核酸溶液A260/A230的消光值比大于或等于2.0；A260/A280大于或等于1.80。A260/A280值过小，说明蛋白未脱净；A260/A230过小，说明有杂质（一般为多酚类或色素）。
这样的提问没有意义
建议自己下去查查资料
仪器性能：拍摄对象：核酸琼脂糖凝胶电泳X光胶片功能：获取并处理图像测定图像光密度操作步骤：1. 开启成像系统，电脑电源2. 打开Quality One，调整光源所示图像3. 观察并调整曝光时间，获得图像并保存4. 调整图像，并对图像进行光密度分析5. 关闭所有电源
一是紫外灯照射出问题了；
也可能是门没关紧，经常接触又断掉，不稳定；
如果成像仪没问题，那很可能就是电脑上操纵拍照的软件在设置上有了问题，应该有配套的说明书的，可以参考；
实在不行，可以报修的，没必要自己找烦恼。
