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主营:研究并生产荧光和发光探针,信号转导研究的试剂
℡ 4000-520-616
℡ 4000-520-616
AAT Bioquest/PhosphoWorks™ Luminometric ATP Assay Kit *Bright Glow*/21610/1 Plate
产品编号:21610
市  场 价:¥42060.00
场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$2103.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/PhosphoWorks™ Luminometric ATP Assay Kit *Bright Glow*/21610/1 Plate
商品介绍
Overview
PrinterFriendlyVersion

Ex/Em(nm)None/560
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategorySmallMoleculeDetection
Anions
RelatedCellCytotoxicity
ProteinKinases
BiochemicalAssays
Adenosinetriphosphate(ATP)playsafundamentalroleincellularenergenics,metabolicregulationandcellularsignaling.ThePhosphoWorks™ATPAssayKitprovidesafast,simpleandhomogeneousluminescenceassayforthedeterminationofcellproliferationandcytotoxicityinmammaliancells.Theassaycanbeperformedinaconvenient96-welland384-wellmicrotiter-plateformat.ThehighsensitivityofthisassaypermitsthedetectionofATPinmanyBIOLOGicalsystems,environmentalsamplesandfoods.ThisPhosphoWorksATPAssayKitcandetectaslowas10cells/well.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,itisrecommendtousewhiteplatestoachievethebestresults.

 

2.PrepareATPassaysolution:

2.1   Thawallthecomponentstoroomtemperaturebeforeuse.

 

2.2   TransferthewholecontentofComponentC(ReactionBuffer,10mL)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:TypicallyATPconcentrationsrangingfrom1nMto1µMareappropriate.

 

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
CitationExplorer

Highthroughputcell-basedassayforidentificationofglycolateoxidaseinhibitorsasapotentialtreatmentforPrimaryHyperoxaluriaType1
Authors:MengqiaoWang,MiaoXu,YanLong,SoniaFargue,NoelSouthall,XinHu,JohnCMcKew,ChristopherJDanpure,WeiZheng
Journal:ScientificReports(2016)

NT1014,anovelbiguanide,inhibitsovariancancergrowthinvitroandinvivo
Authors:LuZhang,JianjunHan,AmandaLJackson,LeslieNClark,JoshuaKilgore,HuiGuo,NickLivingston,KennethBatchelor,YajieYin,TimothyPGilliam
Journal:JournalofHematology&Oncology(2016):91

TheDifferentEffectsofAtorvastatinandPravastatinonCellDeathandPARPActivityinPancreaticNIT-1Cells
Authors:Ya-HuiChen,Yi-ChunChen,Chin-SanLiu,Ming-ChiaHsieh
Journal:JournalofDiabetesResearch(2016)

BPA-inducedDNAhypermethylationofthemastermitochondrialgenePGC-1αcontributestocardiomyopathyinmalerats
Authors:YingJiang,WeiXia,JieYang,YingshuangZhu,HuailongChang,JuanLiu,WenqianHuo,BingXu,XiChen,YuanyuanLi
Journal:Toxicology(2015):21--31

GlutaminepromotesovariancancercellproliferationthroughthemTOR/S6pathway
Authors:LingqinYuan,XiuguiSheng,AdamKWillson,DarioRRoque,JessicaEStine,HuiGuo,HannahMJones,ChunxiaoZhou,VictoriaLBae-Jump
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


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