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当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/PhosphoWorks™ Luminometric ATP Assay Kit *DTT-Free*/21612/1 Plate
商品详细AAT Bioquest/PhosphoWorks™ Luminometric ATP Assay Kit *DTT-Free*/21612/1 Plate
AAT Bioquest/PhosphoWorks™ Luminometric ATP Assay Kit *DTT-Free*/21612/1 Plate
AAT Bioquest/PhosphoWorks™ Luminometric ATP Assay Kit *DTT-Free*/21612/1 Plate
商品编号: 21612
品牌: aatbio
市场价: ¥42060.00
美元价: 25236.00
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)None/560
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategorySmallMoleculeDetection
Anions
RelatedCellSignalingMolecules
BiochemicalAssays
Adenosinetriphosphate(ATP)playsafundamentalroleincellularenergenics,metabolicregulationandcellularsignaling.ThePhosphoWorks™ATPAssayKitprovidesafast,simpleandhomogeneousluminescenceassayforthedeterminationofcellproliferationandcytotoxicityinmammaliancells.Theassaycanbeperformedinaconvenient96-welland384-wellmicrotiter-plateformat.ThehighsensitivityofthisassaypermitsthedetectionofATPinmanyBIOLOGicalsystems,environmentalsamplesandfoods.ThisPhosphoWorksATPAssayKitdoesnotuseDTT,andhasthestableluminescencesignalaslongas4hours.Ithasstableluminescencewithnomixingorseparationsrequired,andformulatedtohaveminimalhands-ontime.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.Preparecells(orsamples):

1.1   Foradherentcells:Platecellsovernightingrowthmediumat1,000-10,000cells/90µL/well(fora96-wellplate)or250-2,000cells/20µL/well(fora384-wellplate).

 

1.2   Fornon-adherentcells:CentrifugethecellsfromtheculturemediumandthensUSPendthecellpelletsinculturemediumat2,000-20,000cells/90µL/wellfora96-wellpoly-Dlysineplateor500-5,000cells/20µL/wellfora384-wellpoly-Dlysineplate.Centrifugetheplateat800rpmfor2minuteswithbrakeoffpriortotheexperiments.

Note1:Eachcelllineshouldbeevaluatedonanindividualbasistodeterminetheoptimalcelldensityforproliferationorcytotoxicityinduction.Fortoxicityassays,startwithmorecells.

Note2:Forallluminescentexperiments,itisrecommendedusingwhiteplatestoachievethebestresults.

 

2.PrepareATPassaysolution:

2.1   Thawallthecomponentstoroomtemperaturebeforeuse.

 

2.2   Transfer10mLofComponentC(ReactionBuffer)intoComponentB(ATPSensor),andmixwell.

 

2.3   Add20µLofComponentA(ATPMonitoringEnzyme)intothesolutionpreparedatStep2.2.

Note:AliquotandstoretheunusedcomponentsAandCat-20oC,andavoidrepeatedfreeze/thawcyclesandpotentialATPcontaminationfromexogenousbiologicalsources.

 

3.RunATPassay:

3.1   Treatcells(orsamples)withtestcompoundsbyadding10µLof10Xcompoundsfora96-wellplateor5µLof5Xcompoundsfora384-wellplateindesiredcompoundbuffer.Forblankwells(mediumwithoutthecells),addthecorrespondingamountofcompoundbuffer.

 

3.2    Incubatethecellplateina37oC,5%CO2incubatorforthedesiredperiodoftime,suchas24,48or96hours.

 

3.3    Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofATPassaysolution(fromStep2.3),andincubateatroomtemperaturefor10-20minutes.

 

3.4   Monitortheluminescenceintensitywithastandardluminometer.

 

4.GenerateastandardATPcalibrationcurve:

Note:AnATPstandardcurveshouldbegeneratedtogetherwiththeaboveassayiftheabsoluteamountofATPinsamplesneedstobecalculated.

 

4.1   MakeaseriesdilutionsofATPinPBSbufferwith0.1%BSAbyincludingasamplewithoutATP(asacontrol)tomeasurebackgroundluminescence.

Note:TypicallyATPconcentrationsrangingfrom0.1nMto1µMareappropriate.

 

4.2   AddthesameamountofthedilutedATPsolutionintoanemptyplate(100µLfora96-wellplateor25µLfora384-wellplate).

 

4.3    Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofATPassaysolution(fromstep2.3).

 

4.4   Incubatethereactionmixtureatroomtemperaturefor10to20minutes.

 

4.5   Recordtheluminescenceintensitywithastandardluminometer.

 

4.6   GeneratetheATPstandardcurve.

References&Citations
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Highthroughputcell-basedassayforidentificationofglycolateoxidaseinhibitorsasapotentialtreatmentforPrimaryHyperoxaluriaType1
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Journal:ScientificReports(2016)

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Journal:JournalofHematology&Oncology(2016):91

TheDifferentEffectsofAtorvastatinandPravastatinonCellDeathandPARPActivityinPancreaticNIT-1Cells
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Journal:JournalofDiabetesResearch(2016)

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Journal:Toxicology(2015):21--31

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Journal:Endocrine-relatedcancer(2015):577--591

JQ1suppressestumorgrowththroughdownregulatingLDHAinovariancancer
Authors:HaifengQiu,AmandaLJackson,JoshuaEKilgore,YanZhong,LeoLi-YingChan,PaolaAGehrig,ChunxiaoZhou,VictoriaLBae-Jump
Journal:Oncotarget(2015):6915

LossofhistonedeacetylaseHdac1disruptsmetabolicprocessesinintestinalepithelialcells
Authors:AlexisGonneaud,NaomieTurgeon,Frančois-MichelBoisvert,FrančoisBoudreau,ClaudeAsselin
Journal:FEBSletters(2015):2776--2783

AneutrophilintrinsicimpairmentaffectingRab27aanddegranulationincysticfibrosisiscorrectedbyCFTRpotentiatortherapy
Authors:KerstinPohl,ElaineHayes,JoanneKeenan,MichaelHenry,PaulaMeleady,KevinMolloy,BakrJundi,DavidABergin,CormacMcCarthy,OliverJMcElvaney
Journal:Blood(2014):999--1009

FluorescencelifetimeimagingunravelsC.trachomatismetabolismanditscrosstalkwiththehostcell
Authors:MártaSzaszák,PhilippSteven,KensukeShima,ReginaOrzekowsky-Schroder,GereonHuttmann,InkeRKonig,WernerSolbach,JanRupp
Journal:PLoSpathogens(2011):e1002108

Gproteincoupledreceptorkinase2interactingprotein1(GIT1)isanovelregulatorofmitochondrialbiogenesisinheart
Authors:JinjiangPang,XiangbinXu,MichaelRGetman,XiShi,StephenLBelmonte,HeidiMichaloski,AmyMohan,BurnsCBlaxall,BradfordCBerk
Journal:Journalofmolecularandcellularcardiology(2011):769--776


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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