| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 330/544 |
| MW | 1123.92 |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | SuperiorLabelingDyes trFluor™DyesandKits |
| Related | LabelingviaThiolGroups BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
SampleProtocolforLabelingProteinswithtrFluor™TbMaleimide:
1. Dissolveyourthiol-containingproteinatconcentration1-10mg/mL(3-10mgistheoptimallabelingconcentration)usingPBSbuffer(20mM,pH7.2).
2. DissolvethetrFluor™TbMaleimideinDMSOattheconcentrationof10-15mg/mL(Forexample,tothe100µgtrFluor™TbMaleimideadd10µLDMSOtomake7mMdyelabelingsolution).
3. MixthetrFluor™TbMaleimide(fromStep2)andproteinsolution(fromStep1)at20:1molarratioofdye/protein,andshakethereactionmixtureatroomtemperaturefor2-4hoursinthedark.
4. Filterthereactionmixturethroughaproteinspincolumnfor100µgto1mgproteinlabelingreaction;orpurifytheconjugateusinggelfiltrationonaproperlysizedSephadexG-25columnifthereactionscaleislargerthan1mg.
5. Collectthedesiredfractionsforyourimmediateuseorfreezedrythemforyourfutureuse.
Note:ThetrFluor™conjugateneedbeusednearneutralpHrange(6.5to7.5).EitheracidicorbasicpHwouldreduceitsfluorescenceintensity.
SampleProtocolforLabelingSmallMoleculeswithtrFluor™TbMaleimide:
1. DissolvetrFluor™TbMaleimide(10-15mg/mL)andyourthiol-containmoleculeinDMSOat1:1.2molarratioofdye/thiol-containmolecule(Forexample,tothe100µgtrFluor™Maleimideadd10µLDMSOtomake7mMdyelabelingsolution).
2. Stirthereactionmixtureatroomtemperaturefor2-4hoursinthedark.
3. PurifytheconjugateusingHPLC(ammoniumacetate/waterandacetonitrile,pH7.0).
4. Collectandpoolthedesiredfractions.5)Combineandfreeze-drythepooledfractions.
Note:ThetrFluor™TbconjugateneedbeusednearneutralpHrange(6.5to7.5).EitheracidicorbasicpHwouldreduceitsfluorescenceintensity.
| References&Citations |  PrinterFriendlyVersion |
1. SavilleL,SpaisC,MasonJL,AlbomMS,MurthyS,MeyerSL,AtorMA,AngelesTS,HustenJ.(2012)Time-ResolvedFluorescenceResonanceEnergyTransferasaVersatileToolintheDevelopmentofHomogeneousCellularKinaseAssays.AssayDrugDevTechnol.
2. LoMC,NgoR,DaiK,LiC,LiangL,LeeJ,EmkeyR,EksterowiczJ,VenturaM,YoungSW,XiaoSH.(2012)Developmentofatime-resolvedfluorescenceresonanceenergytransferassayforcyclin-dependentkinase4andidentificationofitsATP-noncompetitiveinhibitors.AnalBiochem,421,368.
3. PailaYD,KombrabailM,KrishnamoorthyG,ChattopadhyayA.(2011)OligomerizationoftheSEROtonin(1A)receptorinlivecells:atime-resolvedfluorescenceanisotropyapproach.JPhysChemB,115,11439.
4. MartikkalaE,Rozwandowicz-JansenA,HanninenP,Petaja-RepoU,HarmaH.(2011)Ahomogeneoussingle-labeltime-resolvedfluorescencecAMPassay.JBiomolScreen,16,356.
5. GaboritN,LarbouretC,VallagheJ,PeyrussonF,Bascoul-MolleviC,CrapezE,AzriaD,ChardesT,PoulMA,MathisG,BazinH,PelegrinA.(2011)Time-resolvedfluorescenceresonanceenergytransfer(TR-FRET)toanalyzethedisruptionofEGFR/HER2dimers:anewmethodtoevaluatetheefficiencyoftargetedtherapyusingmonoclonalantibodies.JBiolChem,286,11337.
6. LeyrisJP,RouxT,TrinquetE,VerdieP,FehrentzJA,OueslatiN,DouzonS,BourrierE,LamarqueL,GagneD,GalleyrandJC,M"KadmiC,MartinezJ,MaryS,BaneresJL,MarieJ.(2011)Homogeneoustime-resolvedfluorescence-basedassaytoscreenforligandstargetingthegrowthhormonesecretagoguereceptortype1a.AnalBiochem,408,253.
7. PosokhovYO,KyrychenkoA,LadokhinAS.(2010)Steady-stateandtime-resolvedfluorescencequenchingwithtransitionmetalionsasshort-distanceprobesforproteinconformation.AnalBiochem,407,284.
8. Alvarez-CurtoE,WardRJ,PedianiJD,MilliganG.(2010)LigandregulationofthequaternaryorganizationofcellsurfaceM3muscarinicacetylcholinereceptorsanalyzedbyfluorescenceresonanceenergytransfer(FRET)imagingandhomogeneoustime-resolvedFRET.JBiolChem,285,23318.
9. KotaS,ScampaviaL,SpicerT,BeelerAB,TakahashiV,SnyderJK,PorcoJA,HodderP,StrosbergAD.(2010)Atime-resolvedfluorescence-resonanceenergytransferassayforidentifyinginhibitorsofhepatitisCviruscoredimerization.AssayDrugDevTechnol,8,96.
10. VisserAJ,LaptenokSP,VisserNV,vanHoekA,BirchDJ,BrochonJC,BorstJW.(2010)Time-resolvedFRETfluorescencespectroscopyofvisiblefluorescentproteinpairs.EurBiophysJ,39,241.
AAT Bioquest AAT Bioquest是一家位于美国的生物公司，前身为ABD Bioquest，总部位于加利福尼亚州。专门从事光学检测技术十多年，一直致力于光谱学检测领域技术的创新和突破。其独特的光学检测技术，综合了化学、生物学和信息学等各个领域的研究，引领了比色、荧光和发光技术新一代光学探针的浪潮。AAT Bioquest在全球拥有强大的经验丰富的专业分销商网络，为从小型研究机构到《财富》500强企业的各类客户提供卓越的产品和定制服务。
美国AATBioquestInc.(前身是ABDBioquest，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。AATBioquest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。AATBioquest会不断介绍新产品，快速的丰富各个领域的产品。
1）我们提供反应荧光探针和发光探针，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）我们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）我们不断的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）我们致力于开发用于信号转导研究的试剂；5）我们提供生理和神经探针，特别是钙离子指示剂和膜电位探针。
作为AATBioquestInc.的中国区域代理，艾美捷科技为中国客户提供光谱学检测领域，包括吸收（颜色），荧光和发光技术等全系列解决方案。我们也将一如既往更加努力为国内用户提供快捷、方便的高质量产品，同时更为您售前售后全面技术支持。
AATBioquest,Inc.(formerlyABDBioquest,Inc.)develops,manufacturesandmarketsbioanalyticalresearchreagentsandkitstoscientistsengagedinlifesciencesresearch,diagnosticR&Danddrugdiscovery.Wespecializeintheareaofphotometricdetectionsincludingabsorption(color),fluorescenceandluminescencetechnologies.TheCompany"sproductsenablescientistsandbiomedicalresearcherstobetterunderstandbiochemistry,immunology,cellBIOLOGyandmolecularbiology.AATBioquestconstantlyintroducesnewproducts,andoffersarapidlyexpandinglistofproductsthataregroupedintoseveralproductlines.
1)Ourreactivefluorescentandluminescentprobes,biotinsandtagenzymesareusedforlabelingsmalldrugmoleculesandbiopolymers,e.g,proteins,nucleicacidsandcarbohydrates;2)Wedevelopfluorescentandluminescentprobesfordetectingproteins,nucleicacidsandlivecells;3)Weconstantlyintroducenovelfluorescentandluminescentprobesfordetectingvariousenzymes,inparticular,hydrolyticandredoxenzymes;4)Wefocusondevelopingreagentsforsignaltransductionresearch;and5)Wealsoofferphysiologicalandneurologicalprobes,e.g.,calciumindicatorsandmembranepotentialprobes.
Besidesthestandardcatalogproductswealsooffercustomservicetomeetyourspecialresearchneeds.Ourcurrentservicesincludecustomsynthesisofcolorimetric,fluorescentandluminescentprobes,customdevelopmentofbiochemical,cell-basedanddiagnosticassaysandcustomscreeningofyourcompoundlibrariesagainstyourdefinedtargetsusingourvalidatedHTSassays.
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然后是碱性荧光染料，造纸厂用来增加纸的亮度。
直接和分散的荧光黄，印染厂用来增加棉质和化纤面料的艳度
For principle,look at this site:
碘化丙啶(propiolium iodide,PI)能嵌入DNA双螺旋中,可使荧光强度增加约20倍,以488nm波长激发,DNA/PI复合物最大的发射波长约为615nm.
1.小鼠Lewis肺癌细胞DNA含量测定方法
(1).从C57BL/6小鼠上切除肿块,在培养皿内用PBS冲洗.
(2).去除结缔组织及脂肪,剪碎肿块.
(3).小碎片移入1.20×38mm注射针,加压使其通过,于4℃条件下重悬细胞于HBSS中.
(4).将200～300μL细胞悬液(5×105细胞/mL)中加入3mL PI(50μg/mL),染色3LL细胞,于4℃存放20～30分钟.
(5).测定580～750nm之间的发射荧光,以去除末结合PI产生的激发光与发射光谱线之间的重叠部分.
是欢快活泼的光辉色彩，是暖色系中最温暖的色，它使人联想到金色的秋天，丰硕的果实；
是一种富足、快乐而幸福的颜色。让人感觉成一种稳重、含蓄又明快的温暖；
橙色也会带来一种甜甜的感觉。
