Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 501/520 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | NucleicAcidDetection DNADetection |
Related | LabelingCells FluorescenceImaging BiochemicalAssays |
1.Preparing1XAssayBuffer
Preparea1XAssaybufferbydilutingtheconcentratedbuffer20-foldwithsterile,distilled,DNase-freewater.
2.PreparingHelixyteGreen™workingsolution
PrepareHelixyteGreen™workingsolutionbymakinga200-folddilutionoftheconcentratedDMSOsolutionin1Xassaybuffer.Forexample,toprepareenoughworkingsolutiontoassay10samplesina2mLfinalvolume,add50µLofHelixyteGreen™(ComponentA)into10mLofAssayBuffer(fromStep1). Protecttheworkingsolutionfromlightbycoveringitwithfoilorplacingitinthedark.
Note1:Werecommendpreparingthissolutioninaplasticcontainerratherthanglass,asthedyemayadsorbtoglasssurfaces.
Note2:Forbestresults,thissolutionshouldbeusedwithinafewhoursofitspreparation.
3.PrepareserialdilutionsofdsDNAstandard(0to2µg/mL):
3.1 Forhighrangestandardcurve:add30μLof100μg/mLdsDNAstocksolution(ComponentC)to1.47mLof1XAssaybuffer(FromStep1)tohave2000ng/mLdsDNAsolution,andthenperform1:2and1:10serialdilutionstoget1000,100,10,1and0ng/mL.
3.2 Forlowrangestandardcurve:add40μLof2μg/mLdsDNAstocksolution(FromStep3.1)to1.56mLof1XAssaybuffer(FromStep1)tohave50ng/mLdsDNAsolution,andthenperform1:2and1:10serialdilutionstoget25,2.5,0.25,0.025and0ng/mL
4.RundsDNAassay:
4.1 Add1mLofHelixyteGreen™workingsolution(fromStep2)toeachcuvettecontaining1mLofthedsDNAstandard,blankcontrol,andtestsamplestomakethetotaldsDNAassayvolumeof2mL/cuvette.
4.2 Incubatethereactionatroomtemperaturefor5to10minutes,protectedfromlight.
4.3 MonitorthefluorescenceincreasewithaspectroflurometeratEx/Em=490/525nm.
Note:Tominimizephotobleachingeffects,keepthetimeforfluorescencemeasurementconstantforallsamples.
4.4 Thefluorescenceinblankwells(withtheAssaybufferonly)isusedasacontrol,andissubtractedfromthevaluesforthosecuvetteswiththedsDNAreactions.TheDNAconcentrationofthesamplesisdeterminedfromthestandardcurvegeneratedinDNAStandardCurve.
References&Citations | ![]() PrinterFriendlyVersion |
1. PetersonEJ,KireevD,MoonAF,MidonM,JanzenWP,PingoudA,PedersenLC,SingletonSF.(2013)InhibitorsofStreptococcuspneumoniaesurfaceendonucleaseEndAdiscoveredbyhigh-throughputscreeningusingaPicoGreenfluorescenceassay.JBiomolScreen,18,247.
2. ChenY,SonnaertM,RobertsSJ,LuytenFP,SchrootenJ.(2012)ValidationofaPicoGreen-basedDNAquantificationintegratedinanRNAextractionmethodfortwo-dimensionalandthree-dimensionalcellcultures.TissueEngPartCMethods,18,444.
3. MorenoLA,CoxKL.(2010)QuantificationofdsDNAusingtheHitachiF-7000FluorescenceSpectrophotometerandPicoGreendye.JVisExp.
4. DraganAI,Casas-FinetJR,BishopES,StrouseRJ,SchenermanMA,GeddesCD.(2010)CharacterizationofPicoGreeninteractionwithdsDNAandtheoriginofitsfluorescenceenhancementuponbinding.BiophysJ,99,3010.
5. DraganAI,BishopES,Casas-FinetJR,StrouseRJ,SchenermanMA,GeddesCD.(2010)Metal-enhancedPicoGreenfluorescence:applicationtofastandultra-sensitivepg/mlDNAquantitation.JImmunolMethods,362,95.
6. ABIodunOO,GbotoshoGO,AjaiyeobaEO,HappiCT,HoferS,WittlinS,SowunmiA,BrunR,OduolaAM.(2010)ComparisonofSYBRGreenI-,PicoGreen-,and[3H]-hypoxanthine-basedassaysforinvitroantimalarialscreeningofplantsfromNigerianethnomedicine.ParasitolRes,106,933.
7. DraganAI,BishopES,Casas-FinetJR,StrouseRJ,SchenermanMA,GeddesCD.(2010)Metal-enhancedPicoGreenfluorescence:applicationfordouble-strandedDNAquantification.AnalBiochem,396,8.
8. HoldenMJ,HaynesRJ,RabbSA,SatijaN,YangK,BlasicJR,Jr.(2009)FactorsaffectingquantificationoftotalDNAbyUVspectroscopyandPicoGreenfluorescence.JAgricFoodChem,57,7221.
9. IkedaY,IwakiriS,YoshimoriT.(2009)DevelopmentandcharacterizationofanovelhostcellDNAassayusingultra-sensitivefluorescentnucleicacidstain"PicoGreen".JPharmBiomedAnal,49,997.
10. RenX,XuQH.(2009)Label-freeDNAsequencedetectionwithenhancedsensitivityandselectivityusingcationicconjugatedpolymersandPicoGreen.Langmuir,25,43.
AAT Bioquest常用产品报价单
产品名 | 货号 | 公司分类 | 商城分类 | 美金价 |
AAT Bioquest/Fluorescein, disodium salt *CAS 518-47-8*/2/100 mg | 2 | iFluor Rapid Tests | 酸碱缓冲液 | 1088.00 | AAT Bioquest/14-3-3 ζ (Ab-58) Antibody/8B0001/50 ug | 8B0001 | Signaling Intermediate Ab | 重组抗体 | 2828.00 | AAT Bioquest/Opioid Receptor (Phospho-Ser375) Antibody/8A0022/50 ug | 8A0022 | Phospho-Specific Ab | 受体 | 2828.00 | AAT Bioquest/iFluor™ 350 goat anti-mouse IgG (H+L)/16440/200 ug | 16440 | Fluorescent Anti-IgGs | 荧光染料 | 1088.00 | AAT Bioquest/trFluor™ Eu goat anti-mouse IgG (H+L)/16518/100 ug | 16518 | Fluorescent Anti-IgGs | 抗小鼠 | 4278.00 | AAT Bioquest/RPE-streptavidin conjugate/16900/100 ug | 16900 | Streptavidin Conjugates | 其他生物染料 | 1378.00 | AAT Bioquest/GCNT3 Antibody/8C14708/50 ug | 8C14708 | Signaling Intermediate Ab | 功能性抗体 | 2828.00 | AAT Bioquest/Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow*/12518/1 plate | 12518 | Reporter Gene Enzymes | 其它检测试剂盒 | 2103.00 | AAT Bioquest/ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 &mi | 1100 | mFluor™ Dyes and Kits | 标记试剂盒 | 2103.00 | AAT Bioquest/Amplite™ Colorimetric Urea Quantitation Kit *Blue Color*/10058/200 Tests | 10058 | Diagnostic Molecules | 定量试剂盒 | 2828.00 | AAT Bioquest/Screen Quest™ CHO-Gqi Chimera Cell line/38101/Each | 38101 | cAMP GPCR Assays | 常规细胞株 | 0.00 |