Cy5-5-acetate-Sulfo-Cyanine5-5-acetate-DataSheet-生命科学试剂-MCE

Hotline:400-820-3792Inhibitors • ScreeningLibraries • Proteinswww.MedChemECy5.5acetateCat.No.: HY-D0924ASynonyms: Sulfo-Cyanine5.5acetate分子式: C₄₃H₄₈N₂O₁₆S₄分子量: 977.11作用靶点: FluorescentDye作用通路: Others储存方式: PleasestoretheproductundertherecommendedconditionsintheCertificateofAnalysis.BIOLOGICALACTIVITY生物活性 Cy5.5acetate是一种CY染料。CY为花菁(Cyanine)的缩写，是由奇数个甲基单位连接的两个氮原子组成的化合物。菁类化合物具有波长长、吸收和发射可调、消光系数高、水溶性好、合成相对简单等特点。CY系类染料常被用于蛋白，抗体以及小分子化合物的标记，对于蛋白抗体的标记，可以通过简单的混合反应来完成结合，以下我们介绍了蛋白抗体标记的标记方法，具有一定的参考意义。体外研究 Protocol1.ProteinPreparetionInordertoobtainthebestlabelingeffect,pleasepreparetheprotein(antibody)concentrationas2mg/mL.ThepHvalueofproteinsolutionshallbe8.5±0.5.IfthepHislowerthan8.0,1Msodiumbicarbonateshallbeusedforadjustment.Iftheproteinconcentrationislowerthan2mg/mL,thelabelingefficiencywillbegreatlyreduced.Inordertoobtainthebestlabelingefficiency,itisrecommendedthatthefinalproteinconcentrationrangeis2-10mg/mL.Theproteinmustbeinthebufferwithoutprimaryamine(suchasTrisorglycine)andammoniumion,otherwisethelabelingefficiencywillbeaffected.2.DyePreparation(Cy5.5)AddanhydrousDMSOintothevialofCy5.5tomakea10mMstocksolution.Mixwellbypipettingorvortex.3.CalculationofdyedosageTheamountofCy5.5requiredforreactiondependsontheamountofproteintobelabeled,andtheoptimalmolarratioofCy5.5toproteinisabout10.Example:assumingtherequiredmarkerproteinis500μL2mg/mLIgG(MW=150,000),use100μLDMSOdissolve1mgCy5.5,therequiredCy5.5volumeis5.05μL,andthedetailedcalculationprocessisasfollows:1/3 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemE1)mmol(IgG)=mg/mL(IgG)×mL(IgG)/MW(IgG)=2mg/mL×0.5mL/150,000mg/mmol=6.7×10-6mmol2)mmol(Cy5.5)=mmol(IgG)×10=6.7×10-6mmol×10=6.7×10-5mmol3)μL(Cy5.5)=mmol(Cy5.5)×MW(Cy5.5)/mg/μL(Cy5.5)=6.7×10-5mmol×753.88mg/mmol/0.01mg/μL=5.05μL(Cy5.5)4.Runconjugationreaction1)Agoodvolumeoffreshlyprepared10mg/mLCy5.5isslowlyaddedto0.5mLproteinsampleInsolution,gentlyshaketomix,thencentrifugebrieflytocollectthesampleatthebottomofthereactiontube.Don'tmixwelltopreventproteinsamplesfromdenaturationandinactivation.2)Thereactiontubuleswereplacedinadarkplaceandincubatedgentlyatroomtemperaturefor60minutesatintervals.For10-15minutes,gentlyreversethereactiontubulesseveraltimestofullymixthetworeactantsandraisethebarefficiency.5.PurifytheconjugationThefollowingprotocolisanexampleofdye-proteinconjugatepurificationbyusingaSepHadexG-25column.1)PrepareSepHadexG-25columnaccordingtothemanufactureinstruction.2)Loadthereactionmixture(From"Runconjugationreaction")tothetopoftheSepHadexG-25column.3)AddPBS(pH7.2-7.4)assoonasthesamplerunsjustbelowthetopresinsurface.4)AddmorePBS(pH7.2-7.4)tothedesiredsampletocompletethecolumnpurification.Combinethefractionsthatcontainthedesireddye-proteinconjugate.体内研究Cy5.5-labeledfactorVIIaisdevelopedforimaginingcancer.Cy5.5labeledwiththesetargetingproteinsspecificallylocalizetothetumorxenograftsforatleast14daysbutunconjugatedCy5.5doesnotlocalizetoanyxenograftsororgans.Thismethodofimaginganti-tissuefactorinthetumorVECsmaybeusefulindetectingprimarytumorsandmetastasesaswellasmonitoringinvivotherapeuticresponses[1].pH/temperaturesensitivemagneticnanogelsconjugatedwithCy5.5-labledlactoferrin(Cy5.5-Lf-MPNAnanogels)aredevelopedasapromisingcontrastagentforpreoperativeMRIandintraoperativefluorescenceimagingofglioma[2].客户使用本产品发表的科研文献AdvMater.2022Oct4;e2207107.AdvFunctMater.2022:2204636.Small.2022Jul;18(30):e2202337.JNanobiotechnology.2021May26;19(1):156.NanoRes.2023Feb11.Seemorecustomervalidationsonwww.MedChemEREFERENCESZhuS,etal.VisualizingcancerandresponsetotherapyinvivousingCy5.5-labeledfactorVIIaandanti-tissuefactorantibody.JDrugTarget.2015Apr;23(3):257-65.PtaszekM.Rationaldesignoffluorophoresforinvivoapplications.ProgMolBiolTranslSci.2013;113:59-108.2/3 MasterofBioactiveMolecules—您身边的抑制剂大师www.MedChemEShindy,H.A.(2017).Fundamentalsinthechemistryofcyaninedyes:Areview.DyesandPigments,145,505–513.doi:10.1016/j.dyepig.2017.06.029JiangL,etal.pH/temperaturesensitivemagneticnanogelsconjugatedwithCy5.5-labledlactoferrinforMRandfluorescenceimagingofgliomainrats.Biomaterials.2013Oct;34(30):7418-28.LimB,etal.AUniqueRecombinantFluoroprobeTargetingActivatedPlateletsAllowsInVivoDetectionofArterialThrombosisandPulmonaryEmbolismUsingaNovelThree-DimensionalFluorescenceEmissionComputedTomography(FLECT)Technology.The