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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/Amplite™ Fluorimetric Fluorescamine Protein Quantitation Kit *Blue Fluorescence*/11100/200 Tests
产品编号:11100
市  场 价:¥56560.00
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:$2828.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Amplite™ Fluorimetric Fluorescamine Protein Quantitation Kit *Blue Fluorescence*/11100/200 Tests
商品介绍
Overview
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Ex/Em(nm)380/464
MW278.26
CAS#N/A
SolventDMSO
StorageF/D/L
CategoryProteinBiochemistry
Generalproteins
RelatedBiochemicalAssays
Fluorescamineisintrinsicallynon-fluorescentbutreactsrapidlywithprimaryaliphaticamines,includingthoseinpeptidesandproteins,toyieldablue-green-fluorescentderivative.TheAmplite™fluorescamineproteinassaykitprovidesasimplemethodforquantifyingproteinconcentrationinsolutions.ThefluorescentsignalcanbeeasilyreadatEx/Em=380/470nm.ThisAmplite™fluorescamineproteinassaykitcanbeperformedinaconvenient96-wellor384-wellmicrotiter-plateformatandeasilyadaptedtoautomationwithnoseparationstepsrequired.Theassaycanbecompletedwithin30minutes.WiththeAmplite™fluorescamineproteinassaykit,aslittleas3ug/mLofBSAcanbedetected.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.PrepareFluorescamineworkingsolution:AddthewholecontentofDMSO(ComponentB)intothebottleofFluorescamine(ComponentA),andmixwell.

Note:2.5mLoffluorescamineworkingsolutionisenoughfor1plate.Unusedfluorescamineworkingsolutionshouldbestoredinsingleusealiquotsat-20oCandprotectedfromlight.

2.PrepareBSAstandardsolutions:DilutetheappropriateamountofBSAStandard,1mg/mL(ComponentC)intoPBStogetaBSAconcentrationof100-1.56µg/mLwith2Xdilutions.

Table1LayoutofBSAstandardsandtestsamplesinasolidblack96-wellmicroplate

BL

BL

TS

TS

….

….

 

 

 

 

 

 

BS1

CS1

….

….

….

….

 

 

 

 

 

 

BS2

CS2

 

 

 

 

 

 

 

 

 

 

BS3

CS3

 

 

 

 

 

 

 

 

 

 

BS4

CS4

 

 

 

 

 

 

 

 

 

 

BS5

CS5

 

 

 

 

 

 

 

 

 

 

BS6

CS6

 

 

 

 

 

 

 

 

 

 

BS7

CS7

 

 

 

 

 

 

 

 

 

 

               Note:BS=BSAStandards,BL=BlankControl,TS=TestSamples.

               Table2Reagentcompositionforeachwell

BSAStandard

BlankControl

TestSample

SerialDilutions*:75μL

PBS:75μL

75μL

*Note:AddtheserialdilutionsofBSAstandardfrom100to1.56µg/mLwith2XdilutionsintowellsfromBS1toBS7induplicate(seeStep2).

3.Runproteinassay:

3.1   AddtheserialdilutionsofBSAstandardat75μL/well(seeTables1&2)intothe96-wellplate.A0µg/mLBSAcontrolisincludedasblankcontrol.

3.2   Add25μL/welloffluorescamineworkingsolution(fromStep1)intoBSAstandard,blankcontrol,andtestsamples(seeStep2,Table1)tomakethetotalassayvolumeof100µL/well.

Note:Fora384-wellplate,add30μLofsampleand10μLoffluorescamineworkingsolutionintoeachwell.

 

3.3   Incubatethereactionatroomtemperaturefor5to30minutes,protectedfromlight.

3.4   MonitorthefluorescenceincreasewithafluorescenceplatereaderatEx/Em=380/470nm.

References&Citations
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1.   EastwoodD,FernandezC,YoonBY,SheaffCN,WaiCM.(2006)Fluorescenceofaromaticaminesandtheirfluorescaminederivativesfordetectionofexplosivevapors.ApplSpectrosc,60,958.

2.   AdamouR,ColyA,DouabaleSE,SaleckML,Gaye-SeyeMD,TineA.(2005)Fluorimetricdeterminationofhistamineinfishusingmicellarmediaandfluorescamineaslabellingreagent.JFluoresc,15,679.

3.   EggenreichK,ZachE,BeckH,WintersteigerR.(2004)Determinationof4-amino-m-cresoland5-amino-o-cresolbyhighperformanceliquidchromatographyandfluorescencederivatizationusingfluorescamine.JBiochemBiophysMethods,61,35.

4.   SkelleyAM,MathiesRA.(2003)Chiralseparationoffluorescamine-labeledaminoacidsusingmicrofabricatedcapillaryelectrophoresisdevicesforextraterrestrialexploration.JChromatogrA,1021,191.

5.   CesurN,ApakTI,Aboul-EneinHY,OzkirimliS.(2002)LCdeterminationofaminoglutethimideenantiomersasdansylandfluorescaminederivativesintabletformulations.JPharmBiomedAnal,28,487.

6.   BelalF,ABDineH,Al-MajedA,KhalilNY.(2002)Spectrofluorimetricdeterminationofvigabatrinandgabapentininurineanddosageformsthroughderivatizationwithfluorescamine.JPharmBiomedAnal,27,253.

7.   Bantan-PolakT,KassaiM,GrantKB.(2001)Acomparisonoffluorescamineandnaphthalene-2,3-dicarboxaldehydefluorogenicreagentsformicroplate-baseddetectionofaminoacids.AnalBiochem,297,128.

8.   HongC,KondoF.(2000)Detection,quantitation,andidentificationofresidualaminopenicillinsbyhigh-performanceliquidchromatographyafterfluorescaminederivation.JFoodProt,63,1421.

9.   KellyMT,FabreH,PerrettD.(2000)Determinationoftaurineinplasmabycapillaryzoneelectrophoresisfollowingderivatisationwithfluorescamine.Electrophoresis,21,699.

10.   SeguraCarreteroA,Cruces-BlancoC,FernandezPeinadoS,ElBergmiR,FernandezGutierrezA.(1999)Fluorimetricdeterminationofprocaineinpharmaceuticalpreparationsbasedonitsreactionwithfluorescamine.JPharmBiomedAnal,21,969.


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