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当前位置: 首页 > 产品中心 > Fluorescent_dyes > AAT Bioquest/Tide Fluor™8WS马来酰亚胺[TF8WS马来酰亚胺]*近红外发射*/2337/1 mg
商品详细AAT Bioquest/Tide Fluor™8WS马来酰亚胺[TF8WS马来酰亚胺]*近红外发射*/2337/1 mg
AAT Bioquest/Tide Fluor™8WS马来酰亚胺[TF8WS马来酰亚胺]*近红外发射*/2337/1 mg
AAT Bioquest/Tide Fluor™8WS马来酰亚胺[TF8WS马来酰亚胺]*近红外发射*/2337/1 mg
商品编号: 2337
品牌: aatbio
市场价: ¥56560.00
美元价: 33936.00
产地: 美国(厂家直采)
公司:
产品分类: 荧光染料
公司分类: Fluorescent_dyes
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Overview
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Ex/Em(nm)775/807
MW1439.86
CAS#N/A
SolventDMSO
StorageF/D/L
CategorySuperiorLabelingDyes
TideFluor™DyesandKits
RelatedPeptideLabelingReagents
QuencherCPGs
BiochemicalAssays
TideFluor™8WS(TF8WS)familyhasthespectralpropertiessimilartothoseofIRDye800.TheirfluorescenceispH-independentfrompH3to11.ThesecharacteristicsmakethisnewdyefamilymorerobusttopH-sensitiveassays.InsomecasesTF8WS-labeledpeptidesandnucleotidesexhibitstrongerfluorescenceandhigherphotostABIlitythantheoneslabeledwithIRDye800.InpairingwithourTideQuencher™8WS(TQ8WS),avarietyofFRETpeptidesandnucleotidescanbedevelopedfordetectingproteasesandmolecularbeaconswithenhancedsensitivityandstability.ThisTQ8WSproductisusedforpost-labelingofthiol-modifiedoligonucleotidesandpeptidesthatcontaincysteines.
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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

LabelThiol-ModifiedOligonucleotideswithTideFluor™Dyes

Thefollowingprotocolhasbeenoptimizedforlabeling200µg(~6A260nmunits)ofaproprietaryoligonucleotide.Youneedmodifytheprotocoltogetthebestresultsforyourparticularapplicationbymultipleexperimentations.YOURTHIOL-MODIFIEDOLIGOMUSTBETREATEDTOREMOVESMALLTHIOLCOMPOUNDS(SUCHASDTT)THATRAPIDLYREACTSANDCONSUMESDYEMALEIMIDES.

1.      PrepareOligoSolution(SolutionA)

a.      Dissolveyourthiol-modifiedoligo(~200µg)inaPBSbuffer(100µL,pH7.2).

b.     Note1:Theoligonucleotidemustbesynthesizedwithathiolgrouponthe5’end.SeeAppenxidx1forthepurificationofthiolmodifiedoligos.

c.      Note2:Avoidbuffersthatcontainthiolcompounds,suchasDTT,asthesecompeteforconjugationwiththethiol-reactivecompound.

2.      PrepareDyeSolution(SolutionB)

a.      Dissolve1mgdyemaleimidein100µLDMSO(>10mg/mLifpossible)bypipettingupanddown.Centrifugethesolutionstockonthesidesofthevialtothevialbottom.

b.     Note:preparetheDMSOdyesolutionbeforestartingtheconjugation.Extendedstorageofthedyesolutionmayreducethedyeactivity.Anysolutionscontainingthedyeshouldbekeptfromlight.

3.      RunConjugationReaction

a.      Tothedyesolution(B,20-50µL)addtheoligosolution(A,100µL)withstirringorshaking(keepingthereactionmixturefromlight).

b.     Rotateorshakethereactionmixturefor4-6hoursatroomtemperatureonarotatororshaker.

c.      Note:Gentlyvortextapthevialevery10minutesforthefirsthourtoensurethatthereactionsolutionremainswellmixed.Donotmixviolently,asmaterialmaybeleftonthesidesofthevial.Aftersixhours,50–90%ofthethiol-modifiedoligonucleotidemoleculesshouldbelabeled.Thereactionmightbeincubatedovernightifitismoreconvenient.However,overnightincubationwillnotresultinagreaterlabelingefficiencyinmostcases.

4.      PurifyDye-OligoConjugate

a.      Preliminarypurificationbyethanolprecipitationoflabeledoligonucleotide

                                                              i.     Add20µL(one-tenthreactionsolutionvolumeingeneral)of3MNaCland300µLcoldabsoluteethanol(twoandhalfreactionsolutionvolumevolumesingeneral)tothereactionvial.

                                                            ii.     Mixthesolutionwellandplaceitat–20°Cfor30minutes.

                                                           iii.     Centrifugethesolutioninamicrocentrifugeat10,000to15,000×gfor30minutes.

                                                          iv.     Note:Lossofsamplemayoccurifthecentrifugationisnotlongenough.

                                                            v.     Carefullyremovethesupernatant,rinsethepellet1-3timeswithcold70%ethanolanddrybriefly.

                                                          vi.     Note:Someunreactedlabelingreagentmayhaveprecipitatedoverthecourseofthereactionormaybestuckonthewallsofthereactionvial.Thismaterialshouldbecompletelyredissolvedbyextensivevortexmixingbeforecentrifugation.Redissolvingthelabelingreagentensuresthattheprecipitatedoligonucleotidewillbeminimallycontaminatedwithunreactedlabel.

b.     FinalPurificationbyHPLCorbygelelectrophoresis

                                                              i.     SeeAppendixI

 

LabelPeptideswithTideFluor™Dyes

Thefollowingprotocolhasbeenoptimizedforlabeling10mgofaproprietarypeptide(MW~2000)thatcontainsonlyasinglefreethiolgroup.YOUNEEDMODIFYTHEPROTOCOLTOARCHIETHEBESTRESULTSFORYOURPARTICULARAPPLICATIONBYMULTIPLEEXPERIMENTATIONS.

1.      PreparePeptideSolution(SolutionA)

a.      Dissolvethepepetide(tobelabeled)inDMF,DMSOorwater(>10mg/mLifpossible).

b.     Note:Foreffectivelabeling,higherpeptideconcentrationispreferable.

2.      PrepareDyeSolution(SolutionB)

a.      DissolvethedyemaleimideinDMForDMSO(>5mg/mLifpossible).

b.     Note:Foreffectivelabeling,higherdyeconcentrationispreferable.

3.       Mixthepeptideanddyesolutions(fromSteps1and2)inroughlyequalmolarratio.

a.      Note:Eitherpeptideordyemaleimidecanbeusedinexcessdependingonthecostofpeptideanddye.Goodseparationoffreedye,freepeptideanddye-peptideconjugateisanotherimportantfactorthatneedbeconsideredaswelltodecidethateitherpeptideordyeneedbeusedinslightexcess.

4.      Stirthereactionmixtureatroomtemperaturefor12-24hours.

a.       Note:Thisreactionisquiteefficient.Itisgenerallycompletedwithinafewhours.Ifthereactiondoesnotgenerateaconsiderableamountofdesiredconjugatewithin2hours,youneedcheckthereactionpH,averycriticalfactor.TheoptimalpHis4-6.ToolowpHsignificantlydecreasesthereactionratewhileinhighpHsolutionspeptidestendtobeoxidizedtogivethedisulfides.PeptidesoftencontainTFA,thereforeitisimportanttocheckthereactionmixturepHandadjustittopH4-6usingNaHCO3ifnecessary.

5.      PurifyDye-PeptideConjugate

a.      ThereactionsolutionwasconcentratedandpurifiedonaC18columntoaffordthedesiredconjugate.ThefractionswereanalyzedbyHPLC,andthefractionsof>97%puritywerepooledandlyophilized.

b.     Note1:HPLCPurificationConditions:TEABbuffer(triethylammoniumbicarbonate,0.25mmol,pH=7.0-8.0)wasusedasbufferAandacetonitrileasbufferB.TheHPLCwasrunfrom0%Bto30%Bin60min(flowrate:100mL/min).

c.      Note2:Avoidstronglightduringtheoperation.

References&Citations
CitationExplorer

AmechaNISTicmodeltopredicteffectsofcathepsinBandcystatinConβ-amyloidaggregationanddegradation
Authors:TylerJPerlenfein,ReginaMMurphy
Journal:JournalofBIOLOGicalChemistry(2017):jbc--M117

Real-TimeDetectionofaSelf-ReplicatingRNAEnzyme
Authors:CharlesOlea,GeraldFJoyce
Journal:Molecules(2016):1310

DevelopmentofMulti-Parametric/MultimodalSpectroscopyApparatusforCharacterizationofFunctionalInterfaces
Authors:LangZhou,MaryArugula,ChristopherJEasley,CurtisShannon,AleksandrSimonian
Journal:ECSTransactions(2015):9--16

Maternalserumglycosylatedfibronectinasapoint-of-carebioMarkerforassessmentofpreeclampsia
Authors:JuhaRasanen,MatthewJQuinn,AmberLaurie,EricBean,CharlesTRoberts,SrinivasaRNagalla,MichaelGGravett
Journal:Americanjournalofobstetricsandgynecology(2015):82--e1

Arrayofbiodegradablemicroraftsforisolationandimplantationofliving,adherentcells
Authors:YuliWang,ColleenNPhillips,GabrielaSHerrera,ChristopherESims,JenJenYeh,NancyLAllbritton
Journal:RSCadvances(2013):9264--9272

DevelopmentofSNAP-TagFluorogenicProbesforWash-FreeFluorescenceImaging
Authors:XiaoliSun,AihuaZhang,BrendaBaker,LuoSun,AngelaHoward,JohnBuswell,DamienMaurel,AnastasiyaMasharina,KaiJohnsson,ChristopherJNoren
Journal:ChemBioChem(2011):2217--2226

FERRAMENTASPARAESTUDODABIOLOGIADEGPCRS(G-PROTEINCOUPLEDRECEPTORS)
Authors:FredericoMarianettiSoriani,RemoCastroRusso
Journal:Unknown


品牌介绍
美国AAT Bioquest Inc.(前身是ABD Bioquest,Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域,包括吸收(颜色)、荧光和发光技术。AAT Bioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学、免疫学、细胞生物学和分子生物学等领域。AAT Bioquest会不断研发新产品,快速地丰富各个领域的产品。 1)提供反应探针和发光探针,生物素和端粒酶能够应用于标记药物小分子和生物聚合物,如蛋白、核酸以及其他碳水化合物; 2)研究并生产荧光和发光探针用于检测蛋白,核酸和活细胞; 3)不断推出新型的荧光和发光探针用于检测多种酶,特别是检测水解酶和氧化还原酶类; 4)致力于开发用于信号转导研究的试剂; 5)提供生理和神经探针,特别是钙离子指示剂和膜电位探针。
品牌分类
淋巴细胞信号传导 GPCR抗体 羰基活性生物素 细胞代谢 胺反应生物素 链霉亲和素结合物 生物素化抗IgGs 乙酰基特异性抗体 细胞骨架抗体
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