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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™ Ethanol Quantitation Kit/40001/200 Tests
产品编号:40001
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™ Ethanol Quantitation Kit/40001/200 Tests
商品介绍
Overview
PrinterFriendlyVersion

Ex/Em(nm)571/585
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategorySmallMoleculeDetection
DiagnosticMolecules
RelatedBiochemicalAssays
TheABIlitytorapidlyperformquantitativemeasurementsofethanolishighlydesirableinlifescienceresearch,clinicalevaluations,andfoodanddrugindustries.Thisnon-rADIoactiveethanolassayisbasedontheoxidationofethanolbyalcoholoxidase.ThekitusesourAmplite™Redthatmakesthekitrecordableinadualmode,eitherfluorimetric(Ex/Em=540nm/590nm)orcolorimetricreadout(570nm).Thekitprovidesalltheessentialcomponentswithanoptimizedassayprotocol.Theassayisrobust,sensitive,andcanbereadilyadaptedforhighthroughputassaysinawidevarietyofapplicationsthatrequirethemeasurementofethanol.Theassaycanbecompletedwithin30minutes.
SpectrumAdvancedSpectrumViewer

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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Preparestocksolutions:

1.1250XAmplite™EthanolReagentstocksolution:Add40μLofDMSO(ComponentD)intothevialofAmplite™EthanolReagent(ComponentA).Thestocksolutionshouldbeusedpromptly.Anyremainingsolutionshouldbealiquotedandrefrozenat-20oC.

Note1:Avoidrepeatedfreeze-thawcycles.

Note2:TheAmplite™EthanolReagentisunstableinthepresenceofthiolssuchasdithiothreitol(DTT)and2-mercaptoethanol.ThefinalconcentrationofDTTor2-mercaptoethanolinthereactionshouldbenohigherthan10μM.TheAmplite™EthanolReagentisalsounstableathighpH(>8.5).Therefore,thereactionshouldbeperformedatpH7–8.Theprovidedassaybuffer(pH7.4)isrecommended.

1.2100XEthanolEnzymeMix:Add100μLofAssayBuffer(ComponentB)intothevialofEthanolEnzymeMix(ComponentC),andmixwell.

Note:TheunusedEthanolenzymemixsolutionshouldbedividedintosingleusealiquotsandstoredat20oC

2.Prepareassayreactionmixture:

Prepareassayreactionmixtureaccordingtothefollowingtables,protectedfromlight.

Table1Assayreactionmixtureforone96-wellplate

Components

Volume

250XAmplite™ReagentStockSolution(fromStep1.1)

20μL

100XEthanolEnzymeMix(fromStep1.2)

50μL

AssayBuffer(ComponentB)

5mL

Totalvolume

5.07mL

 

Table2Layoutofethanolstandardsandtestsamplesinasolidblack96-wellmicroplate

BL

BL

TS

TS

….

….

 

 

 

 

 

 

ES1

ES1

….

….

….

….

 

 

 

 

 

 

ES2

ES2

 

 

 

 

 

 

 

 

 

 

ES3

ES3

 

 

 

 

 

 

 

 

 

 

ES4

ES4

 

 

 

 

 

 

 

 

 

 

ES5

ES5

 

 

 

 

 

 

 

 

 

 

ES6

ES6

 

 

 

 

 

 

 

 

 

 

ES7

ES7

 

 

 

 

 

 

 

 

 

 

              Note:ES=EthanolStandards,BL=BlankControl,TS=TestSamples.

 

Table3Reagentcompositionforeachwell

EthanolStandard

BlankControl

TestSample

SerialDilutions*:50μL

H2O:50μL

50μL

*Note1:AddtheserialdilutionsofEthanolstandardfrom0.0001%to0.1%intoeachwellfromES1toES7induplicate.

Note2:HighconcentrationofEthanol(e.g.,0.5%,finalconcentration)maycausereducedfluorescencesignalduetotheoveroxidationofAmplite™ethanolreagent(toanon-fluorescentproduct).

3.Runethanolassay:

3.1Prepareanethanolstandardbydilutingtheappropriateamountofthe100%ethanolstandard(ComponentE)intoH2Otoproduceethanolconcentrationrangingfrom0to0.1%,eachinavolumeof50μL.A0%ethanolcontrolisincludedasblankcontrol.Thefinalethanolconcentrationsshouldbetwofoldslower(i.e.,0to0.05%).

3.2Add50μLofassayreactionmixture(fromStep2)intoeachwellofethanolstandard,blankcontrol,andtestsamples(seeStep2,Table3)tomakethetotalethanolassayvolumeof100µL/well.

Note:Fora384-wellplate,add25μLofsampleand25μLofassayreactionmixtureintoeachwell.

3.3Incubatethereactionfor5to30minutesatroomtemperature,protectedfromlight.

3.4MonitorthefluorescenceintensitywithafluorescenceplatereaderatEx/Em=530-570nm/590-600nm(optimalEx/Em=540/590nm),cutoff570nm.

Note:Thecontentsoftheplatecanalsobetransferredtoawhiteclearbottomplateandreadbyanabsorbancemicroplatereaderatthewavelengthof576±5nm.Theabsorptiondetectionhaslowersensitivitycomparedtofluorescencereading.

References&Citations
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DevelopmentofAcyclovir-LoadedAlbuminNanoparticlesandImprovementofAcyclovirPermeationAcrossHumanCornealEpithelialTCells
Authors:PanitaSuwannoi,MullikaChomnawang,NarongSarisuta,StephanReichl,ChristelCMüller-Goymann
Journal:JournalofOcularPharmacologyandTherapeutics(2017)

Enhancementoftheantioxidantandstarchhydrolaseinhibitoryactivitiesofkingcoconutwater(Cocosnuciferavar.aurantiaca)byfermentationwithkombucha‘teafungus’
Authors:MindaniIWatawana,NilakshiJayawardena,ChaminieBGunawardhana,VidurangaYWaisundara
Journal:InternationalJournalofFoodScience&Technology(2016):490--498

Onthedynamicsofkefirvolatome
Authors:Jie-BiHu,SampathGunathilake,Yu-ChieChen,PawelLUrban
Journal:RSCAdvances(2014):28865--28870

pH-Responsivenanoparticlevaccinesfordual-deliveryofantigensandimmunostimulatoryoligonucleotides
Authors:JohnTWilson,SalkaKeller,MatthewJManganiello,ConnieCheng,Chen-ChangLee,ChinonsoOpara,AnthonyConvertine,PatrickSStayton
Journal:ACSnano(2013):3912--3925


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