ProductDescription
AdvancedBiomatrixoffersPhotoGel®RUTKit,apurifiedmethacrylatedgelatinkitforvisIBLelight(400-450nm)photocrosslinkablehydrogels.PhotoGel®provides3Dtunablegelswiththeuniqueattributestobepreparedatvariousconcentrationsandcrosslinkedtoprovidevariousgelstiffness.ThisPhotoGel®kitconsistsofpurifiedmethacrylatedporcinegelatinandaVisibleLightPhotoinitiator(400-450nm).ThegelatinistypeA300bloom.
Table1:
| Item | CatalogNumber | PackageSize | StorageTemperature |
| MethacrylatedGelatin | #5208 | 1gram(2x500mg) | -20°C |
| RutheniumPhotoinitiator | #5246 | 100mg | RoomTemperature |
| SodiumPersulfatePhotoinitiator | #5247 | 500mg | RoomTemperature |
PhotoGel®isproducedfrommethacrylatedgelatinwherethegelatinhasbeenmodifiedbyreactingthefreeamines,primarilytheε-aminesgroupsofthelysineresiduesaswellasthea-aminesgroupsontheN-termini.>75%ofthetotallysineresiduesofthegelatinmoleculehavebeenmethacrylated.ThephotoinitiatorconsistsofRutheniumandSodiumPersulfatetobeformulatedin1XcellculturemediaorPBS,whichallowsvisiblelightphotocrosslinkingofthecollagenat400-450nm.
Forasepticwork,werecommendfilteringthesolubilizedrutheniumandsodiumpersulfatethrough0.2micronbuttonfilters,asthephotoinitiatorsdonotcomesterile.
| Parameter,Testing,andMethod | MethacrylatedGelatin#5208 |
| SterilizationMethod | Filtration |
| Sterility-USPmodified | Nogrowth |
| Form | LyophilizedPowder |
| PackageSize | 1gram(2x500mg) |
| StorageTemperature | -20°C |
| ShelfLife | Minimumof6monthsfromdateofreceipt |
| DegreeofMethacrylation | >75% |
GelatinSource | TypeA,300Bloom,Porcine |
HydrogelYoung"sModulusE(Pa) | Characteristic |
DirectionsforUse
DownloadthefullPDFversionorcontinuereADIngbelow:
3DHydrogelPreparation:
Note:Employasepticpracticestomaintainthesterilityoftheproductthroughoutthepreparationandhandlingofthecollagenandothersolutions.
Note:Thefollowinginstructionsarefora10%gelatinmethacrylatesolution.Recommendedconcentrationsare5-20%.
- Warm10mLofwarm,sterile1XPBSor1Xcellculturemediato>60°C.
- Addthe10mL’sofwarmedsolutiontotheambervialcontaining1gramoflyophilizedgelatinmethacrylate.
- Mixonashakertableorrotatorplateuntilfullysolubilized.Keepwarm(>37°C)ifpossible(eg.placeyourrotatorinanincubator)tohelpwithfullsolubilization.
- Calculatethevolumeofphotoinitiatortoaddbymultiplyingthevolumeofsolubilizedgelatinby0.02.Iftheresultingnumberis200ul,forexample,youwilladd200ulofrutheniumand200ulofsodiumpersulfate.
- Solubilizetherequiredamountofruthenium(perstep4)ataconcentrationof37.4mg/mlin1XPBSorcellculturemedia.
- Solubilizetherequiredamountofsodiumpersulfate(perstep4)ataconcentrationof119mg/mlin1XPBSorcellculturemedia.
- Addtherutheniumtothegelatinsolutionandfullymixuntilsolutionishomogeneous.
- Addthesodiumpersulfatetothegelatin/rutheniumsolutionandmixuntilsolutionishomogeneous.
- Addyourcellstothegelatin/photoinitiatorsolution.
- Dispenseyourgelatin/photoinitiator/cellsolutionintothedesireddish(ie.6-wellplate,48-wellplate).
- Forphotocrosslinking,placeprintedstructuredirectlyundera400-450nmvisiblelightcrosslinkingsource.
Longerexposureallowsmorecrosslinking,thougheachcelltypewithstandsdifferentdegreesoflightandfreeradicals(generatedbythephotoinitiator)thatmediatecrosslinking.
Anyexcessmaterialcanberefrigeratedandstored.Thematerialwillgel.Warmbackupto>30°Cforittobecomeliquidagain.Werecommendonlyaddingphotoinitiatortotheamountofgelatintobeusedatthattime.
ProductApplications
PhotoGel®Gelatinmethacrylatecanbeusedtoformcross-linkedhydrogelsfortissueengineering[1]and3Dprinting.Thecommonformsof3DprintingusingLifeink®300includeextrusion[2][3][4],inkjet[5]andphotolithography[6]).
PhotoGel®hasbeenusedforendothelialcellmorphogenesis,[7]cardiomyocytes,[8]epidermaltissue[9],injectabletissueconstructs[10],bonedifferentiation[11],andcartilageregeneration[12].
Gelatinmethacrylatehasbeenexploredindrugdeliveryapplicationsintheformofmicrospheres[13]andhydrogels[14].
References:
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2878615/
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4040163/
- https://www.ncbi.nlm.nih.gov/pubmed/24112804
- https://www.ncbi.nlm.nih.gov/pubmed/20387987
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5380738/
- https://www.ncbi.nlm.nih.gov/pubmed/28349897/
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3643201/
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4551408/
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4608855/
- http://pubs.acs.org/doi/abs/10.1021/bm401533y
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4252258/
- https://www.ncbi.nlm.nih.gov/pubmed/24590160
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4293288/
- http://www.ijpsonline.com/articles/preparation-and-characterization-of-gelatinpolymethacrylic-acid-interpenetrating-polymeric-network-hydrogels-as-a-phsensitive-deli.html
ProductReferences
ReferencesforPhotoGel®:
Rothrauff,BenjaminB.,etal."EfficacyofThermoresponsive,photocrosslinkablehydrogelsderivedfromdecellularizedtendonandcartilageextracellularmatrixforcartilagetissueengineering."Journaloftissueengineeringandregenerativemedicine12.1(2018):e159-e170.
Rothrauff,BenjaminB.,etal."Anatomicalregion-dependentenhancementof3-dimensionalchondrogenicdifferentiationofhumanmesenchymalstemcellsbysolublemeniscusextracellularmatrix."Actabiomaterialia49(2017):140-151.
Rothrauff,BenjaminB.,GuangYang,andRockyS.Tuan."Tissue-specificbioactivityofsolubletendon-derivedandcartilage-derivedextracellularmatricesonadultmesenchymalstemcells."Stemcellresearch&therapy8.1(2017):133.
Bridge,JackChristopher,etal."Electrospungelatin-basedscaffoldsasanovel3Dplatformtostudythefunctionofcontractilesmoothmusclecellsinvitro."BiomedicalPhysics&EngineeringExpress4.4(2018):045039.
Capella-Monsonís,Héctor,etal."Scaffoldsfortendontissueengineering."HandbookofTissueEngineeringScaffolds:VolumeOne.WoodheadPublishing,2019.259-298.
ProductCertificateofAnalysis
SafetyandDocumentation
SafetyDataSheet
CertificateofOrigin
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。
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原液分装。抗体孵育之前稀释。
做WB的浓度不一定需要很高的。可以设置简单梯度1:200；1:500；1:1000 。看结果再优化
一、广口瓶。
广口瓶是用于盛放固体试剂的玻璃容器，有透明和棕色两种，棕色瓶用于盛放需避光保存的试剂（例如硝酸银等大部分硝酸盐）。广口瓶一般用于存放试剂，瓶口内部磨砂，用于与瓶塞配合使用。
取用试剂时，瓶塞要倒放在桌上，用后将塞塞紧，必要时密封。由于瓶口内侧磨砂，一般跟玻璃磨砂塞配套，因而玻璃塞的广口瓶不能盛放强碱性试剂。如果盛放碱性试剂，要改用橡皮塞，因为强碱与玻璃中的二氧化硅反应，生成硅酸盐，使口与塞粘连。
二、细口瓶。
一种用于存放液体试剂的玻璃容器，细口方便液体倾倒，并且能够避免试剂挥发，因此口比较小。有透明和棕色两种，棕色瓶用于盛放需避光保存的试剂。
取用试剂时，瓶塞要倒放在桌上，用后将塞塞紧，必要时密封。由于瓶口内侧磨砂，一般跟玻璃磨砂塞配套，因而不能盛放强碱性试剂。如果盛放碱性试剂，要改用橡皮塞。
当然做都是可以做，需要带上阴性和阳性的对照组，以区别出是抗体制备的问题还是抗原决定簇被破坏的问题。
