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当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Amplite™ Fluorimetric Total Thiol Quantitation Assay Kit *Green Fluorescence*/5524/200 Tests
商品详细AAT Bioquest/Amplite™ Fluorimetric Total Thiol Quantitation Assay Kit *Green Fluorescence*/5524/200 Tests
AAT Bioquest/Amplite™ Fluorimetric Total Thiol Quantitation Assay Kit *Green Fluorescence*/5524/200 Tests
AAT Bioquest/Amplite™ Fluorimetric Total Thiol Quantitation Assay Kit *Green Fluorescence*/5524/200 Tests
商品编号: 5524
品牌: aatbio
市场价: ¥56560.00
美元价: 33936.00
产地: 美国(厂家直采)
公司:
产品分类: 其它检测试剂盒
公司分类: Other_kits
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍
Overview
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Ex/Em(nm)510/524
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryProteinBiochemistry
Generalproteins
RelatedBiochemicalAssays
Thedetectionandmeasurementoffreethiol(suchasfreecysteine,glutathioneandcysteineresiduesinproteins)isoneoftheessentialtasksforinvestigatingBIOLOGicalprocessesandeventsinmanybiologicalsystems.Themonitoringofreduced(GSH)andoxidizedglutathioneinbiologicalsamplesisessentialforevaluatingtheredoxanddetoxificationstatusofcellsandtissuesinrelationtotheprotectiveroleofglutathioneagainstoxidativeandfree-rADIcal-mediatedcellinjury.Disordersofcysteinemetabolismincludecystinosis,anautosomalrecessivediseaseproducedbyadefectinlysosomaltransport,andcystinuria,acommonheritabledisorderofaminoacidtransport.Therearefewreagentsorassaykitsavailableforquantitatingthiolsinbiologicalsystems.However,allthecommercialkitseitherlacksensitivityorhavetediousprotocols.OurAmplite™FluorimetricTotalThiolQutitationKitprovidesanultrasensitivefluorimetricassayforquantitatingthiolsthatexisteitherinasmallmolecule.Thekitusesaproprietarynon-fluorescentdyethatbecomesstronglyfluorescentuponreactingwiththiol.Thekitprovidesasensitive,one-stepfluorimetricmethodtodetectaslittleas1picomoleofcysteineorGSHina100µLassayvolume(10nMinconcentration).Theassayisrapidandrobust.Itcanbeperformedinaconvenient96-wellor384-wellmicrotiter-plateformatandeasilyadaptedtoautomationwithEx/Em=490nm/520nm.Forrapidquantifyingthiolgroupsinaprotein,werecommendyouuseourkits#5529and#5530thatareoptimizedforquantifyingproteinthiols.
SpectrumAdvancedSpectrumViewer

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Movemouseovergridtodisplaywavelength&intensityvalues.

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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.PrepareGSHstandardstocksolution:

Add200µLofddH2OintotheGSHstandardvial(ComponentC)tomake1mM(1nmol/µL)stocksolution.

Note:TheunusedGSHsolutionshouldbedividedintosingleusealiquotsandstoredat-20oC.

2.Prepare100XThiolite™Greenstocksolution:

Add100µLofDMSO(ComponentD)intothevialofThiolite™Green(ComponentA)tomake100Xstocksolution.

Note:TheunusedThiolite™Greensolutionshouldbedividedintosingleusealiquots,storedat-20oCandkeptfromlight.

3.PrepareGSHreactionmixture:

Add50µLof100XThiolite™Greenstocksolution(fromStep2)into5mLofassaybuffer(ComponentB),andmixwell.

Note1:ThisGSHassaymixture(GAM)isenoughforone96-wellplates.Itisunstableatroomtemperature,andshould 

  beusedpromptlywithin2hoursandavoidexposuretolight.

Note2:Alternatively,onecanmakeGSHAssayMixturebyadding100XThiolite™GreenstocksolutionwithAssay

 Bufferproportionally.

4.PrepareserialdilutionsofGSHstandard(0to10μM):

4.1   Add30μLofGSHstandardstocksolution(fromStep1)to970µLofassaybuffer(ComponentB)togenerate30µM(30pmol/µL)GSHstandard.

Note:DilutedGSHstandardsolutionisunstable.Usewithin4hours.

4.2   Take200μLof30μMGSHstandardsolutiontoperform1:3serialdilutionstoget10,3,1,0.3,0.1,0.03,0.01and0μMserialdilutionsofGSHstandard.

 

4.3   AddGSHstandardsandGSH-containingorotherthiol-containingtestsamplesintoasolidblack96-wellmicroplateasshowninTables1and2.

Note:Treatcellsortissuesamplesasdesired.

 

Table1LayoutofGSHstandardsandtestsamplesinasolidblack96-wellmicroplate

BL

BL

TS

TS

….

….

 

 

 

 

 

 

GS1

GS1

….

….

….

….

 

 

 

 

 

 

GS2

GS2

 

 

 

 

 

 

 

 

 

 

GS3

GS3

 

 

 

 

 

 

 

 

 

 

GS4

GS4

 

 

 

 

 

 

 

 

 

 

GS5

GS5

 

 

 

 

 

 

 

 

 

 

GS6

GS6

 

 

 

 

 

 

 

 

 

 

GS7

GS7

 

 

 

 

 

 

 

 

 

 

Note:GS=GSHStandards,BL=BlankControl,TS=TestSamples.

 

Table2Reagentcompositionforeachwell

GSHStandard

BlankControl

TestSample

SerialDilutions*:50μL

AssayBuffer:50μL

50μL

*Note:AddtheserialdilutionsofGSHstandardfrom0.01μMto10μMintowellsfromGS1toGS7induplicate.

5.RunGSHassay:

5.1   Add50μLofGSHreactionmixture(fromStep3.1)toeachwelloftheGSHstandard,blankcontrol,andtestsamples(seeStep4.3)tomakethetotalGSHassayvolumeof100µL/well.

Note:Fora384-wellplate,add25μLofsampleand25μLofGSHreactionmixtureintoeachwell.

5.2   Incubatethereactionatroomtemperaturefor10minutesto1hour,protectedfromlight.

5.3   MonitorthefluorescenceincreaseatEx/Em=490/520nmwithafluorescenceplatereader.

References&Citations
CitationExplorer

Activationoftranscriptionfactorsinhumanbronchialepithelialcellsexposedtoaqueousextractsofmainstreamcigarettesmokeinvitro
Authors:TakashiSekine,TadashiHirata,ToshikiMine,YasuoFukano
Journal:Toxicologymechanismsandmethods(2016):22--31


品牌介绍
美国AAT Bioquest Inc.(前身是ABD Bioquest,Inc.)是一家为从事生命科学研究、诊断研发及药物开发的科学家研发、生产和销售生物分析研究试剂和试剂盒的公司。公司致力于光谱学检测领域,包括吸收(颜色)、荧光和发光技术。AAT Bioquest的产品帮助全世界的科学家和生物医药研究者更好的了解生物化学、免疫学、细胞生物学和分子生物学等领域。AAT Bioquest会不断研发新产品,快速地丰富各个领域的产品。 1)提供反应探针和发光探针,生物素和端粒酶能够应用于标记药物小分子和生物聚合物,如蛋白、核酸以及其他碳水化合物; 2)研究并生产荧光和发光探针用于检测蛋白,核酸和活细胞; 3)不断推出新型的荧光和发光探针用于检测多种酶,特别是检测水解酶和氧化还原酶类; 4)致力于开发用于信号转导研究的试剂; 5)提供生理和神经探针,特别是钙离子指示剂和膜电位探针。
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淋巴细胞信号传导 GPCR抗体 羰基活性生物素 细胞代谢 胺反应生物素 链霉亲和素结合物 生物素化抗IgGs 乙酰基特异性抗体 细胞骨架抗体
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