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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/FluoroQuest™ Fluorescence Signal Enhancing Solution/20006/5 mL
产品编号:20006
市  场 价:¥27560.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$1378.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/FluoroQuest™ Fluorescence Signal Enhancing Solution/20006/5 mL
商品介绍
Overview
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Ex/Em(nm)None/None
MWN/A
CAS#N/A
SolventN/A
StorageRT
CategoryMicroBIOLOGy
FluorescenceImaging
RelatedBiochemicalAssays
Whenproteinsareconjugatedtofluorescentorganicdyes,fluorescenceemissionofthedyemoleculesisusuallydecreased,sometimesupto50-70%.Thisquenchingphenomenonhasbeenacknowledgedfordecades,butasyet,therearenosimple,practicalmethodstocontrolthefluorescenceofdyesconjugatedtoproteins,especiallyfordyesconjugatedtoimmunoglobulins.WeofferthisFluoroQuest™fluorescencesignalenhancingsolutionthatmightincreasefluorescenceupto2.5-foldincellimagingandflowanalysis.Thisready-to-usesolutionprovidesaneffectivewaytoincreasethesensitivityofdetectionoffluorescentorganiclabelsusedinimmunology,histochemistry,andcellbiology.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.      Platecellsonslidechambersorsterileglasscoverslips,andtreatthecellsasdesired

2.      Performformaldehydefixation.Incubatecellswith3.0–4.0%formaldehydeinPBSatroomtemperaturefor10–30minutes.

 

3.      Washthefixedcells2–3timesinPBS.

Optional:Add0.1%TritonX-100inPBSintofixedcellsfor3to5minutestoincreasepermeABIlity.Rinsethecells2–3timesinPBS.

 

4.      Blockwithblockingagentsuchaswith5%BSAinPBSfor30min.

5.      Diluteprimaryantibodyindilutionbufferasrecommendedinthespecificproduct’sdatasheet.Overlayenoughdilutedantibodytocovercellsoncoversliporeachchamberofthechamberslides.

 

6.      Wash2-3timeswithPBS.

7.      WarmupFluoroQuest™fluorescencesignalenhancingsolutionatroomtemperaturebeforeuse.

8.      DilutefluorescentsecondaryantibodyinFluoroQuest™fluorescencesignalenhancingsolutionandthenstaincellsfor1houratroomtemperature.Generalrangeforsecondaryantibodiesisbetween0.1-10µg/mLforIgGconjugatesformostapplications. Keepslipscoveredorinahumidifiedchambertoavoidevaporation.

 

9.      WashthecellsthreetimeswithPBS.

Note:Additionalstainingwithfluorescentnuclearstainsorphalloidinscanbedoneatthisstep.

10.   Inverteachcoverslipontoacleanedslidewithmountingmedia,preferablyonewithananti-fadepreservative.Sealedgeswithclearpolishifdesired.

11.   Imagingthecellsundermicroscopewithcorrectfilters.Storeslidesinthedarkat4°C.

References&Citations
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1.   CordesT,VogelsangJ,TinnefeldP.(2009)OnthemechanismofTroloxasantIBLinkingandantibleachingreagent.JAmChemSoc,131,5018.

2.   Gallardo-EscarateC,Alvarez-BorregoJ,VonBrandE,DupreE,DelRio-PortillaMA.(2007)RelationshipbetweenDAPI-fluorescencefADIngandnuclearDNAcontent:AnalternativemethodtoDNAquantification?BiolRes,40,29.

3.   WidengrenJ,ChmyrovA,EggelingC,LofdahlPA,SeidelCA.(2007)Strategiestoimprovephotostabilitiesinultrasensitivefluorescencespectroscopy.JPhysChemA,111,429.

4.   DikiciogluE,MeteogluI,OkyayP,CulhaciN,KacarF.(2003)Thereliabilityoflong-termstorageofdirectimmunofluorescentstainingslidesatroomtemperature.JCutanPathol,30,430.

5.   El-RifaiWE,KnuutilaS.(2001)Comparativegenomichybridizationtechnique.MethodsMolMed,50,25.

6.   BerriosM,ConlonKA,ColfleshDE.(1999)Antifadingagentsforconfocalfluorescencemicroscopy.MethodsEnzymol,307,55.

7.   BrockR,HamelersIH,JovinTM.(1999)ComparisonoffixationprotocolsforadherentculturedcellsappliedtoaGFPfusionproteinoftheepidermalgrowthfactorreceptor.Cytometry,35,353.

8.   KolankoCJ,PyleMD,LoatsH,PartonJ,BlakelyWF,NathJ.(1999)Fast-insituhybridizationandimmunoenzymaticcolorpigmentdetectionofmousebonemarrowmicronucleus.BiotechHistochem,74,111.

9.   BenchaibM,DelormeR,PluvinageM,BryonPA,SouchierC.(1996)Evaluationoffivegreenfluorescence-emittingstreptavidin-conjugatedFluorochromesforuseinimmunofluorescencemicroscopy.HistochemCellBiol,106,253.

10.   SwartzDJ,SantiPA.(1996)ImmunofluorescentartifactsduetothepHofantifadingmountingmedia.Biotechniques,20,398.


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