| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 571/585 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | SmallMoleculeDetection DiagnosticMolecules |
| Related | BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
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 | …. | …. |
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ES1 | ES1 | …. | …. | …. | …. |
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ES2 | ES2 |
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ES3 | ES3 |
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ES4 | ES4 |
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ES5 | ES5 |
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ES6 | ES6 |
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ES7 | ES7 |
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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 |  CitationExplorer |
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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1.模板提取（一般为RNA）：Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定：分光光度计或NanoDrop
3.逆转录：逆转录试剂盒（或者一步法试剂盒），这一步可以用普通PCR做，也可以用水域做。
4.荧光定量PCR试剂：通常有用SYBR Green Mix做的，但是这里建议你用EvaGreen做，灵敏度和平行性都要好于SYBR Green，并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他：除了以上的那些还需要离心管、PCR管或板（Axygen反应比较好）、移液枪等，暂时就想到这么多。
1.模板提取（一般为RNA）：Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定：分光光度计或NanoDrop
3.逆转录：逆转录试剂盒（或者一步法试剂盒），这一步可以用普通PCR做，也可以用水域做。
4.荧光定量PCR试剂：通常有用SYBR Green Mix做的，但是这里建议你用EvaGreen做，灵敏度和平行性都要好于SYBR Green，并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他：除了以上的那些还需要离心管、PCR管或板（Axygen反应比较好）、移液枪等，暂时就想到这么多。
MicroBCA试剂测定范围是0.5-20μg/ml
楼主自己看看经销商价格
临床实验室对商品定量试剂盒分析性能的验证.pdf(17431.6k)
如果要是直接注射，就不知道了
