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；
以下产品为AdvancedBioMatrix全球畅销品：1.PureCol 牛源I型胶原蛋白 3mg/ml#5005-100ML2.Nutragen牛源I型胶原蛋白 6mg/ml#5010-50ML3.FibriCol 牛源I型胶原蛋白 10mg/ml#5133-20ML4.VitroCol 人源I型胶原蛋白 #5007-20ML5. 弹性蛋白原 #5052-1MG6.ECMSelectArraykitUltra-36#5170-1EA7.CytoSoft（刚性可变的基底，AdvancedBioMatrix最新添加产品5190-7EA）8. 人III型胶原蛋白 #5021-10MG9. 人IV型胶原蛋白 #5022-5MG10.SilkFibroin溶液 #5154-20ML11.Fibronectin#5080-5MG12.Vitronectin#5051-0.1MG
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凝胶成像仪genegeniusbioimage（syngene公司）系统，我实验室是2006年直接进口了这台仪器，时间太久远，连工程师的号码都是空号了，用过这台仪器的师兄师姐也都毕业不知道了找谁了，请问哪位实验室有这个公司产品的，找一下武汉这边的负责人，我们想联系到再找工程师来维修一下，谢谢！
SIM 凝胶、化学发光图像分析系统为科学家提供了一种新一代36Bit，Bio-1DExpress软件，Windows98/2000/XP均兼容，能够观察分析各种透明或不透明的电泳图像，如EB染色胶，蛋白胶，放射自显影、印迹、斑点印迹等，满足定性，定量分析的迫切需要。
为凝胶图像分析提供了先进，操作简便的解决方法。
为广大从事分子生物学、医院临床检验、法医物证的研究人员提供了一个快捷的解决方式。
它给出更简便、更迅速及更准确的方式以再现、收集、分析图像中的细节。
由图像拍摄、图像处理、数据分析、报告打印等组成一体，符合常规实验的操作思路，做到方便、实用、简单有助于研究人员的正确、迅速地得到凝胶电泳结果照片和分析的其迅速、强大、准确、可靠、经过实践检验，能够使工作更加容易，更加有效。
2．打开电脑，打开并进入成像软件。
ECL拍摄
将拍摄模式切换为“ECL模式”，将滤光轮转到ECL位。选择合适拍摄分辨率（有像素合并功能的机器都有这个功能），点击“启动”。将样品放置在样品台正中间，调整镜头的焦距使样品占据窗口约80%左右，,然后点击“自动曝光”，勾掉“负片”并调整聚焦使预览窗口中的样品图像清晰.（光圈越大，自动曝光所需时间越短。）并先用单帧拍摄，拍摄一张Mark照片。关闭反射白光后给放置在化学发光成像板上的硝酸纤维素膜均匀加上发光液。将拍摄方式设置为“规则积分”，勾上“负片”，并设置的时间和张数，点击“拍摄”按钮即可。
普通凝胶拍摄
1．将拍摄模式切换为“普通模式”，将滤光轮转到UV位（无绿光镜轮的无需调整）。
2．选择合适拍摄分辨率（机器有像素合并功能），点击“启动”。
3. DNA胶拍摄：将DNA胶放置在紫外台正中间，调整焦距使样品占据窗口约80%左右，,然后点击“自动曝光”，并调整聚焦使预览窗口中的样品图像清晰.然后关闭反射白光，开启透射紫外，并微调，确保在紫外下处于清晰状态。
蛋白质胶拍摄：将蛋白质胶放在拆叠白光板的中间，关闭反射白光，开启透射白光，然后点击“自动曝光”，并调整聚焦使预览窗口中的样品图像清晰.。
4.在软件界面点击“拍摄”按钮即可。向左转|向右转
3，预热30分钟
3：获取并处理图像
测定图像光密度
操作步骤，电脑电源
2，获得图像并保存
4：
1,30s后按autoexpose or manualexpose
5. 观察并调整曝光时间. 打开Quality One软件：核酸琼脂糖凝胶电泳
X光胶片
软件功能.调iris. 开启成像系统.打开开关，之后按freeze拍照.xr
4。
BIO-RAD 凝胶成像系统 Universal Hood Ⅱ
仪器性能，保存,focus调节图像清晰度BIO－RAD凝胶电泳成像系统的操作方法
1.按live#47.接通电源和照相机电源
2，文件菜单中gel-doc，并对图像进行光密度分析
5;focus,zoom .打开软件
样品对投射或者反射光有部分的吸收，从而照相所得到的图像上面的样品条带的光密度就会有差异。光密度于样品的浓度或者质量成线性关系。根据未知样品的光密度，通过于已知浓度的样品条带的光密度指相比较就可以得到未知样品的浓度或者质量。这就是图像分析系统定量的基础。采用最新技术的紫外透射光源和白光透射光源使光的分布更加均匀，最大限度的消除了光密度不均造成的对结果的影响。向左转|向右转
暂无品牌问答
