| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 647/670 |
| MW | ~400 |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | EnzymeDetection HorserADIshPeroxidase(HRP) |
| Related |
| Spectrum | AdvancedSpectrumViewer |
1.PrepareAmpliteTMIRworkingsolution:
1.1 Preparea10to25mMstocksolutionofAmpliteTMIRinhigh-quality,anhydrousDMSO.Thestocksolutionshouldbeusedpromptly.Anyunusedsolutionneedtobealiquotedandrefrozenat<-20oC.
Note:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.
1.2 Preparea2XAmpliteTMIRworkingsolution:Onthedayoftheexperiment,eitherdissolveAmpliteTMIRsolidinDMSOorthawanaliquotoftheAmpliteTMIRstocksolutionatroomtemperature.Preparea2Xworkingsolutionof100to250µMin50mMphosphatebufferorbufferofyourchoice,pH7with0.8units/mLperoxidase.AmpliteTMIRfinalconcentrationof50to100µMisrecommendedformeasuringH2O2concentrationinsolution.
Note:AmpliteTMIRisunstableinthepresenceofthiolssuchasDTTandb-mercaptoethanol.Thiolshigherthan10µM(finalconcentration)couldsignificantlydecreasetheassaydynamicrange.NADHandglutathione(reducedfrom:GSH)mayinterferewiththeassay.
2.RunH2O2assayinsupernatants:
2.1 Add50µLof2XAmpliteTMIRworkingsolution(fromStep1.2)intoeachwelloftheH2O2standard,blankcontrol,andtestsamplestomakethetotalH2O2assayvolumeof100µL/well.
Note:Fora384-wellplate,add25µLofsampleand25µLof2XAmpliteTMIRworkingsolutionintoeachwell.
2.2 Incubatethereactionatroomtemperaturefor0to30minutes,protectedfromlight.
2.3 MonitorthefluorescenceincreaseatEx/Em=640/680nmwithafluorescenceplatereader.
Note:Amplite™IRperoxidasesubstrateiseasytobeself-oxidized,soreadthefluorescenceassoonastheH2O2reactionmixtureisaddedtoincreasethesignaltonoiseratio.
2.4 Thefluorescenceinblankwells(withtheassaybufferonly)isusedasacontrol,andissubtractedfromthevaluesforthosewellswiththeH2O2reactions.
3.RunH2O2assayforcells:
Amplite™IRcanbeusedtomeasurethereleaseofH2O2fromcells.Thefollowingisasuggestedprotocolthatcanbemodifiedforyourspecificresearchneeds.
3.1 TheAmpliteTMIRworkingsolutionshouldbepreparedasStep1.2exceptthatthephosphatebuffershouldbereplacedwiththemediathatisusedinthecellculturesystem.Suggestedmediaincluding(a)KrebsRingersPhosphateBuffer(KRPB);(b).HanksBalancedSaltSolution(HBSS);or(c)Serum-freemedia.
3.2 Preparecellsina96-wellplate(50-100µL/well),andactivatethecellsasdesired.
Note:Thenegativecontrols(mediaaloneandnon-activatedcells)areincludedformeasuringbackgroundfluorescence.
3.3 Add50µLofH2O2reactionmixture(fromStep1.2)toeachwellofthecells,andthoseofH2O2standards.
Note:Fora384-wellplate,add25µLofcellsand25µLofH2O2reactionmixtureintoeachwell.
3.4 Incubatethereactionfor0to30minutesatroomtemperature,protectedfromlight.
3.5 MonitorthefluorescenceincreaseatEx/Em=640/680nmwithafluorescenceplatereader.
Note1:Thecontentsoftheplatecanalsobetransferredtoawhiteclearbottomplateandreadbyanabsorbancemicroplatereaderatthewavelengthof670nm.Theabsorptiondetectionhaslowersensitivitycomparedtofluorescencereading.
Note2:Thefluorescencebackgroundincreaseswithtime,thusitisimportanttosubtractthefluorescenceintensityvalueoftheblankwellsforeachdatapoint.
| References&Citations |  CitationExplorer |
AssessmentofTofacitinibandRuxolitinibandtheirAntiInflammatoryEffectsonMyeloperoxidase
Authors:AmberMilton
Journal:(2017)
PatternedPhotonicNitrocelluloseforPseudo-PaperELISA
Authors:JunjieChi,BingbingGao,MiSun,FenglingZhang,EnbenSu,HongLiu,ZhongzeGu
Journal:AnalyticalChemistry(2017)
SpinalCordInflammation:MolecularImagingafterThoracicAorticIschemiaReperfusionInjury
Authors:HassanAlbadawi,JohnWChen,RahmiOklu,YueWu,GregoryWojtkiewicz,BenjaminPulli,JohnDMilner,RichardPCambria,MichaelTWatkins
Journal:Radiology(2016):152222
MyeloperoxidaseNuclearImagingforEpileptogenesis
Authors:YinianZhang,DanielPSeeburg,BenjaminPulli,GregoryRWojtkiewicz,LionelBure,WendyAtkinson,StefanSchob,YoshikoIwamoto,MuhammadAli,WeiZhang
Journal:Radiology(2015):822--830
Myeloperoxidase--Hepatocyte--StellateCellCrossTalkPromotesHepatocyteInjuryandFibrosisinExperimentalNonalcoholicSteatohepatitis
Authors:BenjaminPulli,MuhammadAli,YoshikoIwamoto,MatthiasWGZeller,StefanSchob,JennyJLinnoila,JohnWChen
Journal:Antioxidants&redoxsignaling(2015):1255--1269
Orderedcleavageofmyeloperoxidaseesterbondsreleasesactivesitehemeleadingtoinactivationofmyeloperoxidasebybenzoicacidhydrazideanalogs
Authors:JianshengHuang,ForrestSmith,PeterPanizzi
Journal:Archivesofbiochemistryandbiophysics(2014):74--85
Raisingtheshields:PCRinthepresenceofmetallicsurfacesprotectedbytailor-madecoatings
Authors:FrankDScherag,ThomasBrandstetter,JürgenRühe
Journal:ColloidsandSurfacesB:Biointerfaces(2014):576--582
Measuringmyeloperoxidaseactivityinbiologicalsamples
Authors:BenjaminPulli,MuhammadAli,RezaForghani,StefanSchob,KevinLCHsieh,GregoryWojtkiewicz,JennyJLinnoila,JohnWChen
Journal:PLoSOne(2013):e67976
Micro-volumewall-lessimmunoassaysusingpatternedplanarplates
Authors:KatherineRKozak,JianyongWang,MelvinLye,RashiTakkar,NamyongKim,HyunjaeLee,NooLiJeon,KedanLin,CrystalZhang,WaiLeeTWong
Journal:LabonaChip(2013):1342--1350
Distinguishinginflammationfromtumorandperitumoraledemabymyeloperoxidasemagneticresonanceimaging
Authors:AnneKleijn,JohnWChen,JasonSBuhrman,GregoryRWojtkiewicz,YoshikoIwamoto,MartineLLamfers,AnatOStemmer-Rachamimov,SamuelDRabkin,RalphWeissleder,RobertLMartuza
Journal:ClinicalCancerResearch(2011):4484--4493
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 |
本网站将在规定时间内给予删除等相关处理。
