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当前位置: 首页 > 产品中心 > Other_biological_dyes > AAT Bioquest/FITC-xtra/136/25 mg
商品详细AAT Bioquest/FITC-xtra/136/25 mg
AAT Bioquest/FITC-xtra/136/25 mg
AAT Bioquest/FITC-xtra/136/25 mg
商品编号: 136
品牌: aatbio
市场价: ¥9900.00
美元价: 5940.00
产地: 美国(厂家直采)
公司:
产品分类: 其他生物染料
公司分类: Other_biological_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)496/524
MW620.52
CAS#N/A
SolventDMSO
StorageF/D/L
CategorySuperiorLabelingDyes
iFluor™DyesandKits
RelatedAmine-ReactiveProbes
LabelingviaAminoGroups
AlthoughFITCisstillthemostpopularfluorescentlabelingdyeforpreparinggreenfluorescentbioconjugates,therearecertainlimitationswithFITC,suchasseverephotobleachingformicroscopeimagingandpH-sensitivefluorescence.ProteinconjugatespreparedwithFITC-xtraarefarsuperiorcomparedtothecorrespondingFITCconjugates.FITC-xtraconjugatesaresignificantlybrighterthanFITCconjugatesandaremuchmorephotostable.Additionally,thefluorescenceofFITC-xtraisnotaffectedbypH(4-10).ThispHinsensitivityisamajorimprovementoverFITC,whichemitsitsmaximumfluorescenceonlyatpHabove9.FITC-xtrahasspectralpropertiesalmostidenticalFITC.Inaddition,FITC-xtragivemuchhigherconjugationyieldundermildconjugationconditionsthanFITC.Like5-FITC,FITC-xtraantibodyconjugateshaveexcitationideallysuitedtothe488nmlaserline,makingthemalternativestothecorrespondingFITC-labeledantibodyconjugates.Underthesameconditionstested,FITC-xtraantibodyconjugatesgivemuchhighersignal/backgroundratiosthanthecorrespondingFITC-labeledconjugates.
SpectrumAdvancedSpectrumViewer

Sorry,yourbrowserdoesnotsupportinlineSVG.RelativeIntensity(%)100806040200Sorry,yourbrowserdoesnotsupportinlineSVG.
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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
  1. Prepareproteinstocksolution(SolutionA):
    Mix100µLofareactionbuffer(e.g.,1Msodiumcarbonatesolutionor1MphosphatebufferwithpH~9.0)with900µLofthetargetproteinsolution(e.g.antibody,proteinconcentration>2mg/mlifpossible)togive1mLproteinlabelingstocksolution.

    Note1:ThepHoftheproteinsolution(SolutionA)shouldbe8.5±0.5.IfthepHoftheproteinsolutionislowerthan8.0,adjustthepHtotherangeof8.0-9.0using1Msodiumbicarbonatesolutionor1MpH9.0phosphatebuffer.
    Note2:Theproteinshouldbedissolvedin1Xphosphatebufferedsaline(PBS),pH7.2-7.4.IftheproteinisdissolvedinTrisorglycinebuffer,itmustbedialyzedagainst1XPBS,pH7.2-7.4,toremovefreeaminesorammoniumsalts(suchasammoniumsulfateandammoniumacetate)thatarewidelyusedforproteinprecipitation.
    Note3:ImpureantibodiesorantibodiesstABIlizedwithbovineserumalbumin(BSA)orgelatinwillnotbelabeledwell.Thepresenceofsodiumazideorthimerosalmightalsointerferewiththeconjugationreaction.Sodiumazideorthimerosalcanberemovedbydialysisorspincolumnforoptimallabelingresults.
    Note4:Theconjugationefficiencyissignificantlyreducediftheproteinconcentrationislessthan2mg/mL.Foroptimallabelingefficiencythefinalproteinconcentrationrangeof2-10mg/mLisrecommended.
  2. Preparedyestocksolution(SolutionB):
    AddanhydrousDMSOintothevialofFITC-xtratomakea10-20mMstocksolution.Mixwellbypipettingorvortex.

    Note:Preparethedyestocksolution(SolutionB)beforestartingtheconjugation.Usepromptly.Extendedstorageofthedyestocksolutionmayreducethedyeactivity.SolutionBcanbestoredinfreezerfortwoweekswhenkeptfromlightandmoisture.Avoidfreeze-thawcycles.
  3. Preparethedesiredproteinconjugate:
    Note:Eachproteinrequiresdistinctdye/proteinratio,whichalsodependsonthepropertiesofdyes.Overlabelingofaproteincoulddetrimentallyaffectsitsbindingaffinitywhiletheproteinconjugatesoflowdye/proteinratiogivesreducedsensitivity.Werecommendyouexperimentallydeterminethebestdye/proteinratiobyrepeatingSteps4and5usingaserialdifferentamountoflabelingdyesolutions.Ingeneral4-6dyes/proteinarerecommendedformostofdye-proteinconjugates.
    1. Use10:1molarratioofSolutionB(dye)/SolutionA(protein)asthestartingpoint:Add5µlofthedyestocksolution(SolutionB,assumingthedyestocksolutionis10mM)intothevialoftheproteinsolution(95µlofSolutionA)witheffectiveshaking.Theconcentrationoftheproteinis~0.05mMassumingtheproteinconcentrationis10mg/mLandthemolecularweightoftheproteinis~200KD.
      Note:TheconcentrationoftheDMSOintheproteinsolutionshouldbe<>
    2. Runconjugationreaction.Addtheappropriateamountofdyestocksolution(SolutionB)intothevialoftheproteinsolution(SolutionA)witheffectiveshaking.Continuetorotateorshakethereactionmixtureatroomtemperaturefor30-60minutes.
    3. Repeat#3.2withthemolarratiosofSolutionB/SolutionAat5:1;15:1and20:1respectivelyifdesired.
    4. Purifythedesiredconjugatesusingpremadespincolumnsorothertechniques.
    5. Calculatethedye/proteinratio(DOS)fortheabove4conjugates(seebelow).
References&Citations
CitationExplorer

Roleofreactiveoxygenspecies-mediatedMAPKandNF-$kappa$Bactivationinpolygonatumcyrtonemalectin-inducedapoptosisandautophagyinhumanlungadenocarcinomaA549cells
Authors:TaoLiu,LeiWu,DiWang,HaiyangWang,JinwuChen,ChunlanYang,JinkuBao,ChuanfangWu
Journal:JournalofBiochemistry(2016):mvw040

Determinationofsodiumbenzoateinfoodproductsbyfluorescencepolarizationimmunoassay
Authors:LinlinRen,MengMeng,PengWang,ZhihuanXu,SergeiAEremin,JunhongZhao,YongmeiYin,RimoXi
Journal:Talanta(2014):136--143

ComplexescontainingcationicandanionicpH-sensitiveliposomes:comparativestudyoffactorsinfluencingplasmidDNAgenedeliverytotumors
Authors:YanChen,JiSun,YingLu,ChunTao,JingbinHuang,HeZhang,YuanYu,HaoZou,JingGao,YanqiangZhong
Journal:Internationaljournalofnanomedicine(2013):1573

SwimmingbehaviorofthemonotrichousbacteriumPseudomonasfluorescensSBW25
Authors:LiyanPing,JanBirkenbeil,ShamciMonajembashi
Journal:FEMSmicroBIOLOGyecology(2013):36--44


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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