ProductDescription
Gelin-S®isthiol-modifiedgelatin(denaturedcollagen)andisacomponentoftheHyStem®-C,andHyStem-HPhydrogelkits.MostcellsdonotgrowwellonGelin-S–onlyhydrogels.Instead,Gelin-SshouldbeusedinconjunctionwithGlycosil®(thiol-modifiedhyaluronicacid)orHeprasil®(thiol-modifiedhyaluronicacidwiththiol-modifiedheparin).ReconstitutedGelin-Sremainsliquidat15to37°C.
ThegelatinusedtomakeGelin-SisfromTypeAGelatin,Bloom250,derivedfromporcineskin.
Gelin-S®(thiol-modifiedgelatin)ispackagedin5.0mLvialscontaining50mg.Vialsareblanketedbynitrogenandunderaslightvacuum.
StoreGelin-Sintheoriginalvial,unopened,at-20°Cforuptooneyear.DonotuncaptheGelin-Svialssincetheywillcrosslinkinthepresenceofoxygen.UseasyringeandneedletoaddDGWatertothevials.
Note:Itisrecommendedtoreconstituteeachvialinitsentirety.
DirectionsforUse
Gelin-SispreparedbydissolvingthelyophilizedsolidintheDGWater(oranysterile,degassed,deionizedwater).Whenreconstituted,itwillbein1xphosphatebufferedsaline(PBS),pH~7.4.TheamountofDGWaterusedfordissolutiondependsonthevial.
Gelin-Sshouldbepreparedinthefollowingmanner:
- AllowtheGelin-Svialtocometoroomtemperature.
- Underasepticconditions,usingasyringeandneedle,add1mLofDGwatertothe"1mLGelin-Svial"or5mLofDGwatertothe"5mLGelin-Svial."
- Placethevialhorizontallyonarockerorshaker.Itwilltake<40minutesforthesolidstofullydissolve.Warmingto37°Corlessand/orgentlyvortexingwillspeedupdissolvingtime.Solutionswillbeclearandslightlyviscous.
- Gelin-SwillnotformahydrogelevenifExtralink®isadded.Toformahydrogel,itmustbemixedwithExtralinkandGlycosil®,orHeprasil®.
ProductQ&A
Globularparticleslessthan75kDashouldbeabletofreelydiffusethroughaHyStemhydrogel.
WhenreconstitutedusingDGwater,thepHofeachHyStemcomponentwillbeapproximately7.4-7.6.
Oneyearfromthedateofreceipt,ifstoredproperly.
Anysterile,deionized,degassedwatercanbesubstitutedforreconstitution.However,inordertoensureaccurateandpredictabledissolutionandgelationtimes,ourDGWaterishighlyrecommended,asitisdegassed,blanketedinargon,andhasundergonevalidationtestingwitheachHyStemcomponent.
Gelin-Sprovidescellularattachmentsiteswhenincorporatedinthehydrogel.Gelin-Sisthiol-modified,denaturedcollagenI,derivedfromeitherbovineorporcinesources.Gelin-SisincludedinallHyStem-CandHyStem-HPkits.
Gelin-Shasbeenthiol-modifiedinthesamemannerasthehyaluronaninGlycosil(orHeprasil),sothatitcovalentlycrosslinkswiththeExtralinkintheHyStemhydrogels.
Yes.Peptidesthatcontainacysteineresiduecanbeused.Thecysteineresiduemustbepresentforthepeptidetobecovalentlybondedtothehydrogelsubstrate.
Yes.ECMproteins,suchaslaminin,collagen,fibronectin,orvitronectincanbenon-covalentlyincorporatedintothehydrogelpriortocrosslinking.
HyStemhydrogelsandspongesdifferinhydrationandhomogeneity.HyStemspongesaretypicallypolymerizedhydrogelsthataresubsequentlyfreeze-dried.Theresultingspongeisafibrous,meshnetworkwithporesandnichesthatenablecellstoinfiltrateandadhere.AtrueHyStemhydrogelisanencapsulatingliquidthatpolymerizesaroundsUSPendedcellsinculture.
No.ThecomplianceofthehydrogelsissetbytheamountofExtralinkcrosslinkeradded,theconcentrationofGlycosil(orHeprasil)andGelin-Sused,andtheratioofGlycosil(orHeprasil)toGelin-S.Oncethischemicalstructureofthehydrogelisfixed,itisnotalteredbyprolongedexposuretocellculturemedium.
HyStemspongescanbeterminallysterilizedbyE-beam.HyStemhydrogelshavenotyetbeenvalidatedforusewithE-beamsterilizationmethods.HyStemhydrogelsarenotterminallysterilizedbygammairrADIation.
Gelationtimeisaffectedbymultipleaspectsofthegel’scomposition.Onewaytochangethegelationtimeofahydrogelistovarytheamountofcrosslinkerused.GelswithaloweramountofExtralinkcrosslinkerwillhavealongergelationtimethanthosewithahigheramountofcrosslinker.Changingtheamountofcrosslinkerwillproduceslightchangesingelationtime.GelationtimecanbedramaticallychangedbyvaryingtheGlycosil(orHeprasil)andGelin-Sconcentrations.ConcentratedsolutionsofGlycosil(orHeprasil)andGelin-Swillcreateasolutionwithamuchshortergelationtime.ThiscaneasilybedonebyreconstitutingthecomponentsinasmallervolumeofDGWater.Alternatively,dilutingthesecomponentsinlargervolumesofDGWaterwilldramaticallyincreasethetotaltimetoformthehydrogel.
HyStemHydrogelsarevirtuallytransparentandshouldnotinterferewithmicroscopy.
HyStemhydrogelsmaygeneratemildinflammationaspartofthebody’snaturalhealingprocessinresponsetoinjury.HyStemhydrogelsdonottriggerimmuneresponsewhenusedinvivo.(Theseproductsarenotforhumanuse)
HyStemisdegradedinvivobymatrixmetalloproteinases(Collagenases)andhyaluronidases.
Trypsin,Dipase,collagenase,andhyaluronidasehavebeenusedtohelpdetachcellsfromthesurfaceorfromwithinHyStemhydrogels.
Ingeneral,theporesizeforHyStem-CandHyStem-HPhydrogelsis~17nm.
ProductApplications
Clickonthetitleofthedesiredprotocoltolearnmore:
2DCellGrowthonHyStemHydrogels
HyStem3DCellEncapsulationforCellDeliveryApplicationsGuide
HyStem3DCellEncapsulationinhydrogelsusing96-wellplates
HyStem3DCellEncapsulationinhydrogelsusingTCInserts
EnzymeDigestionofHyStemHydrogelsforRecoveryofEncapsulatedCells
FluorescentLabelingofHyStemHydrogels
CellRecoveryfromSurfaceofHyStemHydrogels
HyStemECMIncorporation
HyStemGelationTimeVariation
HyStemStiffnessVariationProtocolfor7.5mLkit
HyStemStiffnessVariationProtocolfor12.5mLkit
ProductReferences
ReferencesforHyStem®:
Gaetani,R.,etal.(2015)EpicardialapplicationofcardiacProgenitorcellsina3D-printedgelatin/hyaluronicacidpatchpreservescardiacfunctionaftermyocardialinfarction.Biomaterials61:339-348.PMID:17335875.Prestwich,G.D.,etal.(2007)3-Dcultureinsyntheticextracellularmatrices:newtissuemodelsfordrugtoxicologyandcancerdrugdiscovery.AdvEnzymeRegul47:196-207.PMID:17335875.Shu,X.Z.,etal.(2006)Synthesisandevaluationofinjectable,insitucrosslinkablesyntheticextracellularmatricesfortissueengineering.JBiomedMaterResA79:901-912.PMID:16941590.Shu,X.Z.,etal.(2003)Disulfide-crosslinkedhyaluronan-gelatinhydrogelfilms:acovalentmimicoftheextracellularmatrixforinvitrocellgrowth.Biomaterials24:3825-3834.PMID:12818555.
S.Cai,etal.(2005)Injectableglycosaminoglycanhydrogelsforcontrolledreleaseofhumanbasicfibroblastgrowthfactor.Biomaterials,26,6054-6067.D.B.Pike,etal.(2006)Heparin-regulatedreleaseofgrowthfactorsinvitroandangiogenicresponseinvivotoimplantedhyaluronanhydrogelscontainingVEGFandbFGF.Biomaterials,27,5242–5251.G.D.Prestwich,etal.(2007)3-DCultureinSyntheticExtracellularMatrices:NewTissueModelsforDrugToxicologyandCancerDrugDiscovery.invited,Adv.Enz.Res.,inpress(2007).X.Z.Shu,etal,(2006)SynthesisandEvaluationofInjectable,InSituCrosslinkableSyntheticExtracellularMatrices(sECMs)forTissueEngineering.J.BiomedMater.Res.A,79A(4),901-912.
Shu,X.Z.,etal.(2004)Insitucrosslinkablehyaluronanhydrogelsfortissueengineering.Biomaterials25:1339-1348.PMID:14643608.Mehra,T.D.,etal.(2006)Molecularstentingwithacrosslinkedhyaluronanderivativeinhibitscollagengelcontraction.JInvestDermatol126:2202-2209.PMID:16741511.Shu,X.Z.,etal.(2004)AttachmentandspreadingoffibroblastsonanRGDpeptide-modifiedinjectablehyaluronanhydrogel.JBiomedMaterResA68:365-375.PMID:14704979.Ghosh,K.,etal.(2007)CelladaptationtoaphysiologicallyrelevantECMmimicwithdifferentviscoelasticproperties.Biomaterials28:671-679.PMID:17049594.
ProductCertificateofAnalysis
SafetyandDocumentation
CertificateofOrigin
SafetyDataSheet
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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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
据说是见光分解,不过能分解到啥地步就不知道了,至少实验室里的EB都是避光保存了,没听说过那个实验室里把EB放到阳光下晒着。
不过一般接触到皮肤的话如果没有伤口应该没问题(话说我一同学上次一不小心弄了一脸的EB擦干净后就出去晒太阳了.......不过现在貌似没啥事,不知道以后会不会有事...)
但是LZ接触的时候最好还是带上手套,觉得不保险的话就带两层,毕竟那东西是强致癌的。
哈哈不过也不用太害怕,一般实验室在跑电泳的地方都会弄出个EB的污染区,操作时注意下自我的防护,没啥大问题的(话说做实验的有那个是没毒的)。
我想除了PCR方面有些问题外,成像仪的使用和调校我也不是很熟悉,请问各位老师,怎么才能使背景变黑,不至于胶都看得清楚呢?我试过调亮度,GAMMA,但是效果不好.
请教各位,使用成像仪的心得,谢谢了
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
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