Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | None/560 |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | SmallMoleculeDetection Anions |
Related | CellCytotoxicity ProteinKinases BiochemicalAssays |
1.Preparecells(orsamples):
1.1 Foradherentcells:Platecellsovernightingrowthmediumat1,000-10,000cells/90µL/well(fora96-wellplateor250-2,000cells/20µL/wellfora384-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 Transferthewholevialof10mLReactionBuffer(ComponentC)intoATPSensor(ComponentB),andmixwell.
2.3 Add20µLofATPMonitoringEnzyme(ComponentA)intothesolutionpreparedatStep2.2.
Note:AliquotandstoretheunusedComponentsAandCat-20oC.Avoidrepeatedfreeze/thawcyclesandpotentialATPcontaminationfromexogenousbiologicalsources.
3.RunATPassay:
3.1 Treatcells(orsamples)withtestcompoundsbyadding10µLof10Xcompoundsfora96-wellplateor5 µLof5Xcompoundsfora384-wellplateindesiredcompoundbuffer.Forblankwells(mediumwithoutthecells),addthecorrespondingamountofcompoundbuffer.
3.2 Incubatethecellplateina37oC,5%CO2incubatorforadesiredperiodoftime,suchas24,48or96hours.
3.3 Add100µL(96-wellplate)or25µL(384-wellplate)ofATPassaysolution(fromStep2.3)intoeachwell,andincubateatroomtemperaturefor10-20minutes.
3.4 Monitorluminescenceintensitywithastandardluminometer.
4.GenerateastandardATPcalibrationcurve:
Note:AnATPstandardcurveshouldbegeneratedtogetherwiththeaboveassayiftheabsoluteamountofATPinsamplesneedstobecalculated.
4.1 MakeaseriesofdilutionsofATPinPBSbufferwith0.1%BSAbyincludingasamplewithoutATP(asacontrol)formeasuringbackgroundluminescence.
Note:TypicallyATPconcentrationsfrom1nMto10µMareappropriate.
4.2 AddthesameamountofthedilutedATPsolutionintoanemptyplate(100µLfora96-wellplate,and25 µLfora384-wellplate).
4.3 Add100µL/well(96-wellplate)or25µL/well(384-wellplate)ofATPassaysolution(fromStep2.3).
4.4 Incubatethereactionmixtureatroomtemperaturefor10to20minutes.
4.5 Monitortheluminescenceintensitywithastandardluminometer.
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
AAT Bioquest常用产品报价单
产品名 | 货号 | 公司分类 | 商城分类 | 美金价 |
AAT Bioquest/Fluorescein, disodium salt *CAS 518-47-8*/2/100 mg | 2 | iFluor Rapid Tests | 酸碱缓冲液 | 1088.00 | AAT Bioquest/14-3-3 ζ (Ab-58) Antibody/8B0001/50 ug | 8B0001 | Signaling Intermediate Ab | 重组抗体 | 2828.00 | AAT Bioquest/Opioid Receptor (Phospho-Ser375) Antibody/8A0022/50 ug | 8A0022 | Phospho-Specific Ab | 受体 | 2828.00 | AAT Bioquest/iFluor™ 350 goat anti-mouse IgG (H+L)/16440/200 ug | 16440 | Fluorescent Anti-IgGs | 荧光染料 | 1088.00 | AAT Bioquest/trFluor™ Eu goat anti-mouse IgG (H+L)/16518/100 ug | 16518 | Fluorescent Anti-IgGs | 抗小鼠 | 4278.00 | AAT Bioquest/RPE-streptavidin conjugate/16900/100 ug | 16900 | Streptavidin Conjugates | 其他生物染料 | 1378.00 | AAT Bioquest/GCNT3 Antibody/8C14708/50 ug | 8C14708 | Signaling Intermediate Ab | 功能性抗体 | 2828.00 | AAT Bioquest/Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow*/12518/1 plate | 12518 | Reporter Gene Enzymes | 其它检测试剂盒 | 2103.00 | AAT Bioquest/ReadiLink™ Rapid mFluor™ Violet 450 Antibody Labeling Kit *Microscale Optimized for Labeling 50 &mi | 1100 | mFluor™ Dyes and Kits | 标记试剂盒 | 2103.00 | AAT Bioquest/Amplite™ Colorimetric Urea Quantitation Kit *Blue Color*/10058/200 Tests | 10058 | Diagnostic Molecules | 定量试剂盒 | 2828.00 | AAT Bioquest/Screen Quest™ CHO-Gqi Chimera Cell line/38101/Each | 38101 | cAMP GPCR Assays | 常规细胞株 | 0.00 |