ProductDescription
PureCol®CollagenCoated,100X20mmDisheshaveauniformandconsistentapplicationofhighqualityTypeIcollagenonclearpolystyrenesurface.Plateshavearaisedringaroundthecircumference(SureGrip)ofthedishthatprovidesasecuregripofboththedishandlid.Embossedarrowsonthedishandlidprovideorientationforcellsiteobservation.Dishesareasepticallyprocessedandarenon-pyrogenic.Dishesareavailableinquantitiesof10dishespersleeveor50dishespercase.
| Parameter,Testing,andMethod | PureCol®Coated100mmDishes#5028 |
| SterilizationMethod | AsepticallyProcessed |
| Size | 100x20mmDish |
| CollagenUsed | PureCol®TypeI |
| Stackable | Yes |
| VentilationCams | Yes |
| StorageTemperature | 2-30°C |
QuantityperPackage | 10Dishes |
| ShelfLife | Minimumof6monthsfromdateofreceipt |
| FlaskPolymer | Polystyrene |
| TisueCultureTreatedPriortoCoating | Yes |
| GrowthAreaperDish | 58cm2 |
| TypicalWorkingVolumeperDish | 10to20mL |
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
BecausePureCol®hasbeencitedinover2000publications,wehaveonlypostedafewbelow:
Sorensen,JacobR.,etal."Analteredresponseinmacrophagephenotypefollowingdamageinagedhumanskeletalmuscle:implicationsforskeletalmusclerepair."TheFASEBJournal(2019):fj-201900519R.
Sorensen,JacobR.,etal."Analteredresponseinmacrophagephenotypefollowingdamageinagedhumanskeletalmuscle:implicationsforskeletalmusclerepair."TheFASEBJournal(2019):fj-201900519R.
Colaço,E.,etal."HierarchicalCollagen-HydroxyapatiteNanostructuresDesignedThroughLayer-by-LayerAssemblyofCrystal-DecoratedFibrils."J.,HierarchicalCollagen-HydroxyapatiteNanostructuresDesignedThroughLayer-by-LayerAssemblyofCrystal-DecoratedFibrils(May13,2019)(2019).
Schwerdtfeger,LukeA.,etal."Humancolonfunctionexvivo:Dependenceonoxygenandsensitivitytoantibiotic."PloSone14.5(2019):e0217170.
Cardoso,Ana,etal."MiR-144overexpressionasapromisingtherapeuticstrategytoovercomeglioblastomacellinvasivenessandresistancetochemotherapy."Humanmoleculargenetics(2019).
Steele,HannahE.,etal."MechanotransductionofmitochondrialAMPKanditsdistinctroleinflow-inducedbreastcancercellmigration."Biochemicalandbiophysicalresearchcommunications514.2(2019):524-529.
Gehwolf,Renate,etal."GlobalResponsesofIl-1β-Primed3DTendonConstructstoTreatmentwithPulsedElectromagneticFields."Cells8.5(2019):399.
Alexander,Frank,SebastianEggert,andDoriellePrice."Label-FreeMonitoringof3DTissueModelsviaElectricalImpedanceSpectroscopy."(2019):1-24.
Matysik-Woźniak,Anna,etal."ExaminationofKynurenineToxicityonCornealandConjunctivalEpithelium:InvitroandinvivoStudies."Ophthalmicresearch(2019):1-12.
Compton,Clayton,etal."ReconstitutionoftheVentricularEndocardiumWithinAcellularHearts."RegenerativeEngineeringandTranslationalMedicine(2019):1-11.
Müller,A.L.,etal."4.IdentificationofmiR-301ainPrimaryHumanAtrialFibroblastsandBoneMarrow-DerivedMesenchymalProgenitorCellstoAttenuateEndogenousDifferentiationintoPro-FibroticCells."DifferentiationofPrimaryHumanPro-FibroticMesenchymalCellsInfluencedbyExtracellularMatrixEnvironmentDeterminedbyMicro-RNAExpression(2018):130.
Doblinger,Nina,etal."Impactofhydroxyethylstarchandmodifiedfluidgelatinongranulocytephenotypeandfunction."Transfusion(2019).
Elisabeth,etal."Pro-InflammatoryResponsesinHumanBronchialEpithelialCellsInducedbySporesandHyphalFragmentsofCommonDampIndoorMolds."Internationaljournalofenvironmentalresearchandpublichealth16.6(2019):1085.
Dodmane,PuttappaR.,etal."BiphasicchangesinairwayepithelialcellEGFreceptorbindingandphosphorylationinducedbycomponentsofhogbarndust."Experimentallungresearch44.10(2018):443-454.
McClellan,Alyce,etal."Anovelmechanismfortheprotectionofembryonicstemcellderivedtenocytesfrominflammatorycytokineinterleukin1beta."Scientificreports9(2019).
Wang,Weiling,etal."Aquaporin-3deficiencyslowscystenlargementinexperimentalmousemodelsofautosomaldominantpolycystickidneydisease."TheFASEBJournal(2019):fj-201801338RRR.
Teo,JyeYng,etal."SurfacetetheringofstemcellswithH2O2-responsiveanti-oxidizingcolloidalparticlesforprotectionagainstoxidation-induceddeath."Biomaterials201(2019):1-15.
Gehwolf,Renate,etal."3D-EmbeddedCellCulturestoStudyTendonBIOLOGy."(2019):1-11.
ProductCertificateofAnalysis
ProductVideos
SeedingCollagenGelswithCells
Video
30+TypeICollagenOptions
Video
SeeMore
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
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
同学说传代必须是一皿传多皿，一皿传一皿就不算传代~求高手指点~~
传代后5天，依然没有长满，怕影响活力，想传代和冻存，不知这样是否可以？？
想问下各位大神，本人用皿养的PC-9GR细胞突然有大片飘起，也并没有成团状飘起，求解决方法？
各位战友，小弟刚开始养细胞，但是细胞状态一直不好，最近一次传代后，一传二，居然两个皿里细胞状态完全不同，第一个皿还能看见细胞的形态，而第二个皿几乎看不见贴壁的细胞，请问大家遇见过这种情况吗？
做细胞实验快半年了，一直都还挺顺的，这次五一放完假回来，复苏一支HK2，操作都跟以前一样，没想到出了好多问题：
冻存方法：包裹棉花直接-80℃过夜，第二天转移到液氮
冻存时间：1个半月前冻存的细胞，密度保证没问题，冻存前状态也好
复苏方法：液氮取出后37℃水浴，约2分钟溶解，加入6倍体积的完全培养基，800转离心5分钟，弃去上清，1ml完全培养基重悬，转移入培养皿（进口一次性塑料培养大皿），补足完全培养基，培养箱培养
第二天看细胞全都没贴壁，但是也没死，聚集成团装飘着，没有污染。不想重新离心加重机械损伤，就一直试试看的心态放在培养箱里养着了。又重新复苏一支，还是一样的结果，全飘着没贴壁。不死心，就往前面复苏的那一皿里直接加了1ml的血清，相当于18%的血清比例，过了一天去看，这下细胞都贴壁了。没有另外添加血清的那一皿就还是没贴壁。
另外还有一支以前复苏的HK2，也是一样的方法复苏的，那次复苏很好，细胞基本没什么死的，也都贴壁了，养在皿里状态也不错，但是拿来铺板就还是不贴壁，同样的培基（10%血清），铺板就一个不贴，皿里就都可以贴上。
求助各位战友：
1、复苏不贴壁是为什么？
2、增加血清比例能使复苏的细胞贴壁，这样的细胞是不是可以认为状态并不好，以后的培养是不是要一直这么高比例的血清？还是可以培养一段时间逐步减少血清比例？
3、仍然是10%血清，为什么铺板就不贴壁，而皿里的就没事？是不是铺板的时候也要增加血清比例呢？那在板里干预的过程中是不是要一直保持高比例的血清培养？
4、我的冻存及复苏方法是否有错？我觉得我的HK2从形态、生长速度上来说状态应该是不错的，而且我已经更换了全新的培基、血清和双抗，重新配置了完全培养基，不知道为什么会出现这种不贴壁的问题
拜托各位集思广益，细胞实验已经为了这个不贴壁的问题停滞快2周，心急如焚啊，拜托各位！
实验需要请问有没有养THP-1细胞的同仁？能否寄一皿我，我在广州南方医科大学，不胜感激！
如题~
已经基本养好了贴壁的原代细胞了，准备鉴定了，看到好多都在说细胞爬片的问题，我可以不做爬片直接在皿里操作吗？具体需要注意些什么呢？感谢各位大神
它最初由在德国生物学家罗伯特·科赫手下工作的细菌学家朱利斯·理查德·佩特里（1852－1921）于1887年设计，故又称为“佩特里皿”。
====================
其实就是常见的圆形的上下盖培养皿，你可以看看它的英文，就叫做petri dish，直译过来就叫做——佩特里小碟子——也就是皮氏培养皿
=====================
第二，三段为原创手打……第一段为摘抄
