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当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Cell Meter™ Fluorimetric Cell Cytotoxicity Assay Kit/22781/1000 Tests
商品详细AAT Bioquest/Cell Meter™ Fluorimetric Cell Cytotoxicity Assay Kit/22781/1000 Tests
AAT Bioquest/Cell Meter™ Fluorimetric Cell Cytotoxicity Assay Kit/22781/1000 Tests
AAT Bioquest/Cell Meter™ Fluorimetric Cell Cytotoxicity Assay Kit/22781/1000 Tests
商品编号: 22781
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)571/585
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategoryCellAnalysis
CellCytotoxicity
RelatedApoptosisandCytotoxicity
CellApoptosis
BiochemicalAssays
OurCellMeter™assaykitsareasetoftoolsformonitoringcellviABIlity.Thereareavarietyofparametersthatcanbeusedformonitoringcellviability.Themeasurementofmitochondrialdehydrogenases(e.g.LDH)activityisawell-acceptedassaytoquantifycellnumbersandmonitorcellviability.Thiscellviabilityassaykitprovidesafast,simple,accurateandhomogeneousassayforthefluorometricdetectionofviablecells.Thisassayisbasedontheobservationthatblueandnon-fluorescentresazurinisreducedtoapinkfluorescentdye(resorufin)byacceptingtheelectronfrommitochondrialrespiratorychaininlivecells.Theamountofresorufinproducedisdirectlyproportionaltothenumberoflivingcells.ThedetectionsensitivityofcellproliferationandcytotoxicityassaysusingthiskitishigherthanotherassayssuchasMTT.Sincethekitcomponentsarequitestablewithminimalcytotoxicity,alongerincubation(suchas24to48hours)ispossIBLe.Theassaycanbeperformedinaconvenient96-welland384-wellmicrotiter-plateformat.Thecharacteristicsofitshighsensitivity(<100 cho="" cells),="" non-radioactive="" and="" no-wash="" method="" made="" the="" kit="" suitable="" for="" high="" throughput="" screening="" of="" cell="" proliferation="" or="" cytotoxicity="" against="" a="" variety="" of="" compounds="" and="" adaptable="" for="" a="" wide="" variety="" of="" instrument="" platforms.="" the="" kit="" provides="" all="" the="" essential="" components="" with="" an="" optimized="" assay="" protocol.="" it="" is="" suitable="" for="" proliferating="" and="" non-proliferating="" cells,="" and="" can="" be="" used="" for="" both="" suspension="" and="" adherent="" cells.="" using="" 20="" ul="" of="" reagents="" per="" well="" in="" a="" 96-well="" format,="" this="" kit="" provides="" sufficient="" reagents="" to="" perform="" 1000="" assays.="" using="" 10="" ul="" of="" reagents="" per="" well="" in="" a="" 384-well="" format,="" this="" kit="" provides="" sufficient="" reagents="" to="" perform="" 2,000="" assays.="">
SpectrumAdvancedSpectrumViewer

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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Preparecellsandtestcompounds:

1.1   Plate100to10,000cellsperwellinatissueculturemicroplatewithblackwallandclearbottom.Addtestcompoundsintothecells,andincubateforadesiredperiodoftime(suchas24,48or96hours)ina37°C,5%CO2incubator.Forblankwells(mediumwithoutthecells),addthesameamountofcompoundbuffer.Thesuggestedtotalvolumeis100µLfora96-wellplate,and50µLfora384-wellplate.

 

1.2   Setupthefollowingcontrolsatthesametime.

Positivecontrolcontainscellsandknownproliferationorcytotoxicityinducer.

Negativecontrolcontainscellsbutnotestcompounds.

Vehiclecontrolcontainscellsandthevehicleusedtodelivertestcompounds.

Non-cellcontrolcontainsgrowthmediumwithoutcells.

Note:LDHinserumwillcontributetobackgroundfluorescence.

Testcompoundcontrolcontainsthevehicleusedtodelivertestcompounds[Hank’sbalancesaltsolution(HBSS)orphosphate-bufferedsaline(PBS)]andtestcompound.Sometestcompoundshavestrongautofluorescenceandmaygivefalsepositiveresults.

Note:Matchthetotalvolumeofallthecontrolsto100µLfora96-wellplateor50µLfora384-wellplatewithgrowthmedium.

 

2.Assayprocedures:

2.1   ThawandwarmuptheAssaySolution(ComponentA)to37°C,andmixitthoroughlybeforestartingtheexperiments.

 

2.2   Add20µL/well(96-wellplate)or10µL/well(384-wellplate)ofAssaySolution(ComponentA).Mixthereagentsbyshakingtheplategentlyfor30seconds.

 

2.3    Incubatethecellsina37oC,5%CO2incubatorfor1-24hours,protectedfromlight.

Note1:Theappropriateincubationtimedependsonthemetabolismrateoftheindividualcelltypeandcellconcentrationused.Optimizetheincubationtimeforeachexperiment.

Note2:Extremelyprolongedincubationtimeisnotrecommendedsinceresazurincouldbeconvertedtocolorlessdihydroresorufin.

 

2.4   Monitorthefluorescenceintensity(bottomread)atEx/Em=540/590nm.Alternatively,readtheO.D.at570nm(thereferencewavelengthshouldbe600nm)todeterminethecellviabilityineachwell.

 

3.Performdataanalysis:

3.1   ThebackgroundfluorescencereADIngfromthenon-cellcontrolwellissubtractedfromthevaluesforthosewellscontainingthecells.

Note:Thebackgroundfluorescenceoftheblankwellsmayvarydependingonthesourcesofthegrowthmediaorthemicrotiterplates.

 

3.2   Thefluorescencereadingineachwellindicatesthecellnumberinthatwell.

 

3.3   Calculatethepercentageofcellviabilityforsamplesandcontrolsbasedonthefollowingformula:

 

%Cellviability=100×(Fsample-Fo)/(Fctrl-Fo)

Fsampleisthefluorescencereadinginthepresenceofthetestcompound.

Fctrlisthefluorescencereadingintheabsenceofthetestcompound(vehiclecontrol).

Foistheaveragedbackground(non-cellcontrol)fluorescenceintensity.

References&Citations
PrinterFriendlyVersion

1.   McNichollBP,McGrathJW,QuinnJP.(2006)Developmentandapplicationofaresazurin-basedbiomassactivitytestforactivatedsludgeplantmanagement.WaterRes.

2.   OriowoMO.(2006)Afluorometricstudyofrelativeocularlenscytosensitivitytomultipurposecontactlenssolutionsusingtheresazurinassaymethod.ToxicolInVitro,20,1548.

3.   UmubyeyiAN,MartinA,ZissisG,StruelensM,KaritaE,PortaelsF.(2006)EvaluationoftheresazurinmicrotiterassayforrapiddetectionofofloxacinresistanceinM.tuberculosis.IntJTubercLungDis,10,808.

4.   NatecheF,MartinA,BarakaS,PalominoJC,KhaledS,PortaelsF.(2006)ApplicationoftheresazurinmicrotitreassayfordetectionofmultidrugresistanceinMycobacteriumtuberculosisinAlgiers.JMedMicrobiol,55,857.

5.   CobanAY,CekicCihanC,BilginK,UzunM,AkgunesA,CetinkayaE,DurupinarB.(2006)RapidsusceptibilitytestforMycobacteriumtuberculosistoisoniazidandrifampinwithresazurinmethodinscrew-captubes.JChemother,18,140.

6.   ZrimsekP,KosecM,KuncJ,MrkunJ.(2006)Determinationofthediagnosticvalueoftheresazurinreductionassayforevaluatingboarsemenbyreceiveroperatingcharacteristicanalysis.AsianJAndrol,8,343.

7.   RolonM,VegaC,EscarioJA,Gomez-BarrioA.(2006)DevelopmentofresazurinmicrotiterassayfordrugsensibilitytestingofTrypanosomacruziepimastigotes.ParasitolRes,99,103.

8.   TizzardAC,BergsmaJH,Lloyd-JonesG.(2006)Aresazurin-basedbiosensorfororganicpollutants.BiosensBioelectron,22,759.

9.   Abu-AmeroKK,BosleyTM.(2005)Detectionofmitochondrialrespiratorydysfunctionincirculatinglymphocytesusingresazurin.ArchPatholLabMed,129,1295.

10.   Anoopkumar-DukieS,CareyJB,ConereT,O"SullivanE,vanPeltFN,AllshireA.(2005)Resazurinassayofradiationresponseinculturedcells.BrJRadiol,78,945.


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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。
品牌分类
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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