ProductOverview
TheTRA-1-81monoclonalantibodyreactswithapluripotent-stem-cell-specificantigenexpresseduponthesurfaceofhumanembryonicstem(ES)cells,embyronalcarcinoma(EC)cells,andembryonicgerm(EG)cells.TheexpressionofTRA-1-81antigenisstage-specific;ashumanECandEScellsundergodifferentiation,expressionofTRA-1-81antigenislost,makingitawidelyusedMarkertocharacterizehumanEScellsandtomonitortheirdifferentiation.TRA-1-81antibodyrecognizesacarbohydrateepitopeexpressedonpodocalyxin,asialoglycoproteinstructurallyrelatedtoCD34.Podocalyxinisatransmembraneglycoproteinwhichhasbeenimplicatedinthedevelopmentofaggressivenessinavarietyofcancers,includingbreastandprostatecancer.
ProductSpecifications
Size
100μl
Concentration
0.5mg/ml
Clone
Tra-1-81
Isotype
MouseIgM,κ
Immunogen
Humanembryonalcarcinomacellline2102Ep
Reactivity
Human
Preparation
Thisantibodywaspurifiedbyaffinitychromatography.
Formulation
Phosphate-bufferedsolution,pH7.2,and0.09%sodiumazide
StorageandStABIlity
Storeat4˚Cprotectedfromlight.Donotfreeze.Stablefor6monthsfromdateofreceiptwhenstoredasdirected.
QualityControl
Testedbyimmunocytochemistryandflowcytometrytoensureproductquality.
RecommendedUsage
Thesuggesteduseofthisantibodyisa1:100dilutionforimmunocytochemistryand0.25µgper1x106viablecellsin100µlforflowcytometry.Forapplicationspecificprotocols,pleasereferenceProtocol:ImmunocytochemistryandProtocol:FlowCytometryonlineatwww.Stemgent.com/support/protocols.
SpecificationSheets
- 09-0011SpecificationsSheet
SafetyDataSheets
- 09-0011SafetyDataSheet
Protocols
- Protocol:ImmunoflourescenceStainingofCellsforFlowCytometry
- Protocol:Immunocytochemistry
RelatedProducts
- mRNAReprogrammingKit(00-0071)
- microRNABoosterKit(00-0073)
References
- Andrews,P.W.,Banting,G.,Damjanov,I.,Arnaud,D.,andAvner,P.(1984)Threemonoclonalantibodiesdefiningdistinctdifferentiationantigensassociatedwithdifferenthighmolecularweightpolypeptidesonthesurfaceofhumanembryonalcarcinomacells.Hybridoma3:347-361.
- Yu,J.,Vodyanik,M.A.,Smuga-Otto,K.,Antosiewicz-Bourget,J.,Frane,J.L.,Tian,S.,Nie,J.,Jonsdottir,G.A.,Ruotti,V.,Stewart,R.,Slukvin,I.I.,andThomson,J.A.(2007)Inducedpluripotentstemcelllinesderivedfromhumansomaticcells.Science318:1917-1920.
- Henderson,J.K.,Draper,J.S.,Baillie,H.S.,Fishel,S.,Thomson,J.A.,Moore,H.,andAndrews,P.W.(2002)Preimplantationhumanembryosandembryonicstemcellsshowcomparableexpressionofstage-specificembryonicantigens.StemCells20:329-337.
- Schopperle,W.M.,andDeWolf,W.C.(2007)TheTRA-1-60andTRA-1-81humanpluripotentstemcellmarkersareexpressedonpodocalyxininembryonalcarcinoma.StemCells25:723-730.
AdditionalPublications
- PoleganovMA;EminliS;BeissertT;HerzS;MoonJ-I;GoldmannJ;BeyerA;HeckR;BurkhartI;RoldanDB;TureciO;YiK;HamiltonB;SahinU."Efficientreprogrammingofhumanfibroblastsandblood-derivedendothelialProgenitorcellsusingnonmodifiedRNAforreprogrammingandimmuneevasion."HumanGeneTherapy26:751(2015)
- PaullD;SevillaA;ZhouH;HahnAK;KimH;NapolitanoC;TsankovA;ShangL;KrumholzK;JagaseesanP;WoodardCM;SunB;VilbouxT;ZimmerM;ForeroE;MoroziewiczDN;MartinezH;MalicdanMCV;WeissKA;VensandLB;DusenberryCR;PolusH;SyKTL;KahlerDJ;GahlWA;SolomonSL;ChangS;MeissnerA;EgganK;NoggleSA."Automated,high-throughputderivation,characterizationanddifferentiationofinducedpluripotentstemcells."NaureMethods12:885(2015)
- Wang,Y-C.,Nakagawa,M.,Garitaonandia,I.,Slavin,I.,Altun,G.,Lacharite,R.M.,Nazor,K.L.,Tran,H.T.,Lynch,C.,L.,Leonardo,T.R.,Liu,Y.,Peterson,S.E.,Laurent,L.,C.,Yamanaka,S.,Loring,J.F.(2011)Specificlectinbiomarkersforisolationofhumanpluripotentstemcellsidentifiedthrougharray-basedglycomicanalysis.CellRes.21(11):1551-1563.
- Nagy,K.,Sung,H-K.,Zhang,P.,Laflamme,S.,Vincent,P,Agha-MohammADI,S.,Woltjen,K.,Monetti,C.,Michael,I.P.,Smith,L.C.,Nagy,A.InducedPluripotentStemCellLinesDerivefromEquineFibroblasts.StemCellReviewsandReports7:693(2011).
- Mack,A.,Kroboth,S.,Rajesh,D.,Wang,W.B.GenerationofInducedPluripotentStemCellsfromCD34+CellsacrossBloodDrawnfromMultipleDonorswithNon-IntegratingEpisomalVectors.PLoSOne;6(11):e27956.(2011)
Stemgent mRNA体细胞重编程系统
产品特点
1.快速 Fast
Stemgent® MicroRNA-Enhanced mRNA Reprogramming System只需2周即可将靶细胞重编程为iPS细胞，与仙台病毒（Sendai Virus）、非整合型DNA载体（Episomal DNA Vectors）等传统iPS重编程方法相比，缩短实验周期超过50%以上（Figure. 1）。
Figure 1. Stemgent® mRNA重编程系统、仙台病毒依赖的重编程系统以及游离型DNA载体依赖的重编程系统重编程时程比较。
简便 Easy
与病毒、DNA载体依赖的iPS重编程手段不同，使用Stemgent® MicroRNA-Enhanced mRNA Reprogramming System对靶细胞进行重编程，生成的iPS细胞无需进行复杂耗时的下游筛选程序，只需简简单单6个步骤，即可轻松构建iPS细胞系。
| Description | Day | |
| Step 1 | Material Preparation | Prior to starting |
| Step 2 | Plate Cells | 0 |
| Step 3 | Transfect Cells | 1 to 12 |
| Step 4 | Identify Cells | 13-14 |
| Step 5 | Pick and Passage iPS Cell Colonies | 15-16 |
| Step 6 | Maintain iPS Cell Cultures | 16+ |
注：因所用细胞类型及实验条件不同，上述时间轴可能略有不同，仅供参考。
Figure 2. Stemgent® MicroRNA-Enhanced mRNA Reprogramming System重编程过程时间表、显微镜下观察到的重编程过程中靶细胞形态变化。第12天，形成的iPS克隆表达多能型Marker TRA-1-81。
高效 Efficient
病毒、DNA载体等依赖的iPS重编程技术，其重编程效率介于0.00001%到1%之间不等，而Stemgent® MicroRNA-Enhanced mRNA Reprogramming System的重编程效率远远大于1%，极大地提高了iPS克隆的产出率。
表1. 不同类型iPS重编程手段效率比较
| Reprogramming Method | Efficiency | Integrating | Screening |
| RNA | >1% | No | No |
| 仙台病毒（Sendai virus） | 0.01-1% | No | Yes |
| DNA载体（Episomal/Minicircle） | 0.00% | Possible | Yes |
| 慢病毒（Lentivirus） | 0.001-0.01% | Yes | Yes |
| 腺病毒（Adenovirus） | 0.0001-0.001% | Possible | Yes |
| Protein | 0.00% | No | No |
安全 Safe
某些iPS研究领域，例如iPS临床研究、治疗应用等，对iPS细胞的安全性提出了极高的要求。使用Stemgent® MicroRNA-Enhanced mRNA Reprogramming System重编程靶细胞，无需担心构建的iPS细胞系可能含有病毒载体残留，亦无需担心无关的基因整合进宿主基因组，是目前适用于临床研究及治疗的最理想选择。
而以仙台病毒（Sendai Virus）为载体的iPS重编程手段，其病毒基因组虽然不会整合进宿主染色体，仙台病毒本身也不具备复制能力，但是为了构建真正意义上的virus-free的iPS细胞系，通常需要经过一个10-20代、耗时费力的筛选过程，以确保最终得到的iPS细胞系无病毒载体残留。以慢病毒（Lentivirus）体的iPS重编程手段，病毒基因组能够永久整合进宿主基因组，会给后续的iPS细胞分析带来干扰。
高性价比 Cost Effective
以仙台病毒为载体的iPS重编程试剂盒非常昂贵，综合重编程效率、时间及劳动力成本、安全性等多重考虑，Stemgent® MicroRNA-Enhanced mRNA Reprogramming System无疑是性价比的上乘之选。
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