ProductDescription
AdvancedBiomatrixoffersPhotoHA®,amethacrylatedhyaluronicacid(HA)forphotocrosslinkablehydrogels.Thesehydrogelsprovidenative-like3DHAgelswiththeuniqueattributestobepreparedatvariousconcentrationsandphotocrosslinkedtoprovidevariousgelstiffness.Thekitcomeswith100mgoflyophilizedmethacrylatedhyaluronicacidand100mgofLAPphotoinitiator(405nmbluelightphotocrosslinking).
| Item | CatalogNumber | PackageSize | StorageTemperature |
| MethacrylatedHyaluronicAcid | #5212 | 100mg | -20°C |
| LAPPhotoinitiator | #5269 | 100mg | 2-10°C |
Hyaluronicacidisthemostabundantglycosaminoglycaninthebodybeinganimportantcomponentofseveraltissuesthroughoutthebody.Whileitisabundantinextracellularmatrices,hyaluronicacidalsocontributestotissuehydrodynamics,movementandproliferationofcells,andparticipatesinanumberofcellsurfacereceptorinteractions.
Forthemajorityofcelltypes,itisrecommendedtoaddadditionalECMproteinstothehyaluronicacidhydrogels.Theproteinsprovideimportantcellbindingsites.
Storage:
Theproductshipsonfrozengelpacks.Uponreceipt,storethePhotoHA®at-20°C.StoretheLAPat2to10°C.Theproductandcomponentsarestableforaminimumof1yearatreceiptinpowderform.
Oncesolubilized,thePhotoHA®canbestoredat2-10°Cfor1month.Thephotoinitiatorcanbestoredfornomorethan2weeksoncesolubilized.
| Parameter,Testing,andMethod | MethacrylatedHyaluronicAcid#5212 |
| SterilizationMethod | Filtration |
| Sterility-USPmodified | Nogrowth |
| Form | LyophilizedPowder |
| PackageSize | 100mg |
| StorageTemperature | -20°C |
| ShelfLife | Minimumof6monthsfromdateofreceipt |
| ShelfLifeAfterReconstitution | 1Month |
| DegreeofMethacrylation | 45-65% |
| MolecularWeight | 100-150kDa |
NMRAnalysis | Characteristic |
HydrogelYoung"sModulusE(Pa) | Characteristic |
SwellingCharacteristicsofPhotoHA®:
50μLhydrogelsfabricatedin4.7mmdiametermoldswereimagedbeforeandafterincubationinphosphatebufferedsalineat25°Cfor24hours.ThediameterofhydrogelswerequantifiedusingImageJsoftware.Statisticalcomparisonsbetweengroups(n=3)wereperformedviaone-wayANOVAwithposthoctestingandsignificancedeterminedatp<0.05.
CompressiveModulusofPhotoHA®:
Dynamicmechanicalanalysis(Q800,TAInstruments)wasperformedon50μLhydrogelsfabricatedin4.7mmdiametermolds.Hydrogelsweresecuredwithinafluidcupviaa0.01Npre-loadandcompressedto30%strainatarateof0.5Nmin-1.TheYoung’smodulusofeachhydrogelwascalculatedastheslopeofgeneratedstress-straincurvesbetween10%and20%strain.StatisticalcomparisonbetweenMeHAconcentrations(n=3)wasperformedviaStudentst-testwithtwo-tailedcriteriaandsignificancedeterminedatp<0.05.
ReactionBehaviorofPhotoHA®:
Rheologicaltimesweeps(AR2000stresscontrolledrheometer,TAInstruments;0.5%strain,1Hz,25°C)ofMeHAcrosslinkingwithexposuretoUVlight(=320-390nm)andinthepresenceof0.05wt%Irgacure2959(I2959).After1minute,themacromersolution(i.e.MeHAandI2959)wasexposedtoUVlight,resultingintheplateauofmodulibefore5minutes.
DirectionsforUse
DownloadthefullDirectionsforUsePDForcontinuereADIngbelow:
3DHyrdogelPreparation:
Note:Employasepticpracticestomaintainthesterilityoftheproductthroughoutthepreparationandhandlingofthecollagenandothersolutions.
Recommendedconcentrationsare5-30mg/ml(0.5-3.0%).
Note:Thefollowingrecommendedinstructionsarefora1%hyaluronicacid(HA)methacrylatesolution.Adjustmentstothisprotocolmayberequiredforvariousconcentrations.
- Add10mlof1Xphosphatebuffersaline(PBS),waterorcellculturemediatothe100mgoflyophilizedmethacrylatedHApowder.
- Mixonashakertableorrotatorplateuntilfullysolubilized(~30to60minutes)at2-10°C.Note:SolubilizationtimesmayvarydependingonthedesiredconcentrationandvolumeofPBS,waterormediumadded,
- Calculatethevolumeofphotoinitiatortoaddbymultiplyingthevolumeofsolubilizedhyaluronicacidby0.02.Iftheresultingnumberis200ul,forexample,youwilladd200ulofLAP.
- SolubilizetherequiredamountofLAP(perstep3)ataconcentrationof17mg/mlin1XPBSorcellculturemedia.
- AddthecalculatedvolumeofphotoinitiatortotherequiredvolumeofHAmethacrylatesolutionandmixthoroughly.
- AddyourcellstotheHAmethacrylate/photoinitiatorsolution.
- DispenseyourHAmethacrylate/photoinitiator/cellsolutionintothedesiredcultureware(i.e.6-wellplate,48-wellplate).
- Forphotocrosslinking,placethehydrogelsolutiondirectlyundera405nmlightcrosslinkingsource.
ProductQ&A
Yes.TherearequiteafewpublicationscitingvarioushyaluronidaseprotocolsfordigestingPhotoHAhydrogels.Hereareafew:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5460858/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4840832/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4986518/
TheHAweusecomesfrombiotechnologicalproduction,wherethehyaluronicacidisextractedfromthecellwallsofthebacteriaStreptococcuszooepidemicus.
ProductCellAssay
Humanmesenchymalstemcells(20x106/mL)wereencapsulatedin50μLhydrogels(~4.7mmx2mm).Hydrogels(1wt%PhotoHA®)werefabricatedwith0.05wt%Irgacure2959andexposureto2mW/cm2light(320-390nm)for10minutes.After24hours,encapsulatedcellswerestainedwithcalceinAMandethidiumhomodimerandsubsequentlyimagedonaLeicaSP5confocalmicroscope(usingFITC/TRITCsequentialscans).
ProductApplications
PhotoHA®MethacrylatedHyaluronicacidcanbeusedtoformhydrogelsforex-vivoengineeringofautologouscartilagetissue[1]orasamesenchymalstemcellcarrierincartilagerepair[2].
BecausethestiffnesscanbewidelyadjustedbyalteringconcentrationorUV-lightexposure,methacrylatedHAhasbeenusedtomeasuretheeffectsofmatrixstiffnessoncellphenotypeandfunction[3][4].
MethacrylatedHAcanbeusedfor3Dbioprinting(extrusion[5],inkjet[5]andphotolithographic[6])tocreatestructuresthatpromoteosteogenicdifferentiationofMSC’s[7].
ThehightunABIlityofhyaluronicacidmethacrylateallowsittobemixedwith,andreinforceothertypesofhydrogels(suchascollagen,orgelatinmethacrylate)[8].
References:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5717235/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627486/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5541838/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5447944/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5615317/
https://www.ncbi.nlm.nih.gov/pubmed/21773726/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5460858/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5748291/
ProductReferences
ReferencesforPhotoHA®:
Khetan,Sudhir,andOwenCorey."Maintenanceofstemcellviabilityanddifferentiationpotentialfollowingcryopreservationwithin3-dimensionalhyaluronicacidhydrogels."CryoBIOLOGy(2019).
Poldervaart,M.T.etal.3Dbioprintingofmethacrylatedhyaluronicacid(MeHA)hydrogelwithintrinsicosteogenicity.PlosOne12,(2017).
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。
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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一、免疫组化试剂盒一般包括：
1、特异性的一抗
2、免疫组化检测系统。
3、有的同时还具备显色系统。
二、不同的公司内容有差别，所以购买时一定了解清楚：
1、一抗是做人的还是兔、鼠的组织；
2、检测系统是ABC/SP/非生物素/其他；
3、试剂盒的内容都有什么？有没有显色系统？
4、如果没有，自己根据前面的检测系统需配哪个显色系统。（其他的辅助试剂如PBS缓冲液、抗原修复液等一般另卖或自配）
三、免疫组化抗体即指特异性一抗，是用来标记的指标，免疫组化过程的其他试剂一律另外配备。
另外：做实验时，如遇出结果困难，记得做阳性片对照，敢于怀疑检测系统或显色系统的问题。
用双氧水去除过氧化物酶时，我是自己配的3%双氧水滴到片上而非将片浸入双氧水，这样可否？
用博士德的DAB染色，配制方法为：1mlH20中加入DAB、H2O2、TBS各一滴，我配制的时候发现将DAB加入H20时很容易沉淀，要震荡才能混匀，且将配好的DAB滴到玻片上时，DAB会结为很小块的微颗粒悬浮在液体中但并不染色，是否DAB有问题？
现在打算换中衫的试剂盒，代理商说中衫的试剂盒包括了二抗和DAB，想请教一下该试剂盒的具体内容有些什么？
万分感谢！
上海好多的供应商的，具体还是看你选择了，其实试剂盒产品都差不多，什么远慕生物，古朵生物...好多的额
