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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谢谢谢
只要引物设计的可以在基因组上做,那么理论上是没问题的!
只需要用RT-PCR荧光定量试剂盒里面的第二个,专门用来做Real-timePCR的那些试剂就可以了!
如果要是直接注射,就不知道了
试剂盒里有详细的说明书,告诉你样品需要多少量,每个试剂需要加入多少量,和详细的实验步骤,一般买来就可以用,不用人教。
所以你问一个样需要多少量是没法回答的,测定过程是要加很多种试剂的。
定性ELISA只能粗略表示样本待测物含量在某个特定值以上或以下,定性ELISA通常设置阳性对照(P)、和阴性对照(N),结果分别用“阴性”和“阳性”表示。ELISA定性测定的“阴性”和“阳性”的判断标准是试剂盒所确定的阳性判断值,即cut-off值。
定量ELISA是通过一系列不同已知浓度的标准品所对应的OD值做出标准曲线,然后将样本的OD值代入标准曲线,计算出样本中待测物的含量。百特纯大分子Meretciel的定量ELISA试剂盒还可以。
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