![AAT Bioquest/Tide Quencher™ 2WS acid [TQ2WS acid]/2050/25 mg](images/AAT201712/hq-figure-1-for-tide-quencher-2ws-acid-tq2ws-acid.png)
| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 512/None |
| MW | 490.60 |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | LabelingQuenchers TideQuencher™Dyes |
| Related | PeptideLabelingReagents Amine-ReactiveProbes BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
LabelAmino-ModifiedOligonucleotideswithTideQuencher™Dyes
Thefollowingprotocolhasbeenoptimizedforlabeling200µg(~6A260nmunits)ofaproprietaryoligonucleotide.Youneedmodifytheprotocoltogetthebestresultsforyourparticularapplicationbymultipleexperimentations.YOURAMINO-MODIFIEDOLIGOMUSTBETREATEDTOREMOVEAMMONIATHATRAPIDLYREACTSANDCONSUMESDYESUCCINIMIDYLESTERS.
1. PrepareOligoSolution(SolutionA)Dissolveyouramino-modifiedoligo(~200µg)inatetraboratebuffer(100µL,pH8.5±0.5).
a. Note1:Theoligonucleotidemustbesynthesizedwithanaminegrouponthe5’end.SeeAppenxidx1forthepurificationofamino-modifiedoligos.
b. Note2:Avoidbuffersthatcontainprimaryamines,suchasTris,asthesecompeteforconjugationwiththeamine-reactivecompound.
2. PrepareDyeSolution(SolutionB)
a. Dissolve1mgdyeSEin100µLDMSO(>10mg/mLifpossible)bypipettingupanddown.Centrifugethesolutionstockonthesidesofthevialtothevialbottom.
b. Note:preparetheDMSOdyesolutionbeforestartingtheconjugation.Extendedstorageofthedyesolutionmayreducethedyeactivity.Anysolutionscontainingthedyeshouldbekeptfromlight.WedonotrecommendthatyoustoretheDMSOdyesolutionforfutureuse.
3. RunConjugationReaction
a. Tothedyesolution(B,20-50µL)addtheoligosolution(A,100µL)withstirringorshaking(keepingthereactionmixturefromlight).
b. Rotateorshakethereactionmixturefor4-6hoursatroomtemperatureonarotatororshaker.
c. Note:Gentlyvortextapthevialevery10minutesforthefirsthourtoensurethatthereactionsolutionremainswellmixed.Donotmixviolently,asmaterialmaybeleftonthesidesofthevial.Aftersixhours,50–90%oftheamine-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
LabelPeptideswithTideQuencher™Dyes
Thefollowingprotocolhasbeenoptimizedforlabeling10mgofaproprietarypeptide(MW~2000)thatcontainsonlyasinglefreeaminogroup.YOUNEEDMODIFYTHEPROTOCOLTOARCHIETHEBESTRESULTSFORYOURPARTICULARAPPLICATIONBYMULTIPLEEXPERIMENTATIONS.
1. PreparePeptideSolution(SolutionA)
a. Dissolveyourpeptide(~10mg)inDMF(~1ml).
b. Note1:Thepeptidemustbeneutralizedwithabasesuchastriethylamineorpotassiumcarbonate.
c. Note2:Avoidbuffersthatcontainprimaryamines,suchasTris,asthesecompeteforconjugationwiththeamine-reactivecompound.
2. PrepareDyeSolution(SolutionB)
a. Dissolve5mgdyeSEin500µLDMF(>10mg/mLifpossible)bypipettingupanddown.
b. Note:preparetheDMFdyesolutionbeforestartingtheconjugation.Extendedstorageofthedyesolutionmayreducethedyeactivity.Anysolutionscontainingthedyeshouldbekeptfromlight.WedonotrecommendthatyoustoretheDMFdyesolutionforfutureuse.
3. RunConjugationReaction
a. Tothedyesolution(B,500µL)addthepeptidesolution(A,1mL)withstirringorshaking(keepingthereactionmixturefromlight).
b. Stirthereactionmixturefor4-6hoursatroomtemperature.
4. 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
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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分子生物学实验的染料主要涉及到核酸染料和蛋白质染料.核酸染料主要有EB（溴化乙锭,高致癌性）,goldview,sybr green（实时定量PCR时常用染料）.这些染料可以和核酸双链分子特异性结合发出强荧光而被检测到.蛋白质染料最常用的是考马斯亮蓝 R-250,硝酸银.其中硝酸银有时也用于核酸染色.
染料分为天然染料和人工染料两种。天然染料有胭脂虫红、地衣素、石蕊和苏木素等，它们多从植物体中提取得到，其成分复杂，有些至今还未搞清楚。目前主要采用人工染料，也称煤焦油染料，多从煤焦油中提取获得，是苯的衍生物。多数染料为带色的有机酸或碱类，难溶于水，而易溶于有机溶剂中。为使它们易溶于水，通常制成盐类。
染料可按其电离后染料离子所带电荷的性质，分为酸性染料、碱性染料、中性（复合）染料和单纯染料四大类。 标本干燥后即进行固定，固定的目的有三个：
1）杀死微生物，固定细胞结构。
2）保证菌体能更牢的粘附在载玻片上，防止标本被水冲洗掉。
3）改变染料对细胞的通透性，因为死的原生质比活的原生质易于染色。
健那绿——高中唯一一个活体染色剂。染线粒体的，染成蓝绿色
产品主要应用：点击化学(Clickchemistry)、蛋白质组学研究中的双向荧光差异凝胶电泳(2DDIGE)和实时荧光定量PCR(RealtimePCR)。
氨基类染料是包含自由氨基的活性染料，染料可与活化羧酸衍生物和其他亲电子的试剂结合。比如：氨基与EDC-活化的羧基结合。
相关产品如下：
中文名英文名产品编号分子结构Cy7.5胺Cy7.5amineAGF1350A[img]/KindEditor_4.0.1/attached/image/20130704/20130704110804_5250.jpg[/img]Cy5胺Cy5amineAGF1332A[img]/KindEditor_4.0.1/attached/image/20130704/20130704110715_7750.jpg[/img]相似系列产品：
抗体、核酸、蛋白质等生物分子标记染料
羰基活性荧光染料
巯基反应性染料
羧酸类染料
请各位大侠给予帮助！！
谢谢！！
而且样品中的无水硫酸钠未变色,而做标准曲线的五个和空白对照的变为蓝色了,请高手指教,多谢!
还有,是否变蓝对测定结果有影响吗?
谢了哈
欢迎你！请下次规范发贴:)
2、苏丹三 脂肪 橙红
3、苏丹四 脂肪 红
4、双缩脲 蛋白质 紫
5、龙胆紫 染色质 紫
6、碘 淀粉 蓝
7、健那绿 线粒体 绿
8、甲基绿 DNA 绿
9、吡罗红 RNA 红
10、溴麝香草酚蓝 CO2 由蓝变绿再变黄
11、重铬酸钾 酒精 酸性条件下由橙色变成灰绿
12、醋酸洋红（龙胆紫、改良苯酚品红） 染色质 红
13、台盼蓝 检验活死细胞 死细胞会被染成蓝色（不常用）
取片剂一片，照溶出度测定法（中国药典2000年版二部附录ⅩC第三法），以水250ml为溶剂，转速为每分钟50转，依法操作，分别经15、30、45、60、75、210分钟取溶液滤过，精密量取续滤液5ml于分液漏斗中，加入pH7.4磷酸盐缓冲液5ml，5ml0.3%溴麝香草酚蓝溶液，用15ml氯仿分两次萃取，合并萃取液，加入0.4g无水硫酸钠，照分光光度法（中国药典2000年版二部附录ⅥA），在410nm波长处测得溶出A值。现15、30、45、60、75、210分钟溶出A值分别为0.0413、0.0544、0.0437、0.0479、0.0394、0.0302。（测定吸收度偏小是否不准，有影响。）
请各位站友指教。
