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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/OxiVision Green™ hydrogen peroxide sensor/21505/1 mg
产品编号:21505
市  场 价:¥27560.00
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:$1378.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/OxiVision Green™ hydrogen peroxide sensor/21505/1 mg
商品介绍
Overview
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Ex/Em(nm)490/514
MW~600
CAS#N/A
SolventDMSO
StorageF/D/L
CategoryNeuroBIOLOGy
ReactiveOxygenSpecies
RelatedCellFunctionalAnalysis
RedoxEnzymes
BiochemicalAssays
DespitetheimportanceofH2O2tohumanhealthanddisease,themolecularmechanismsofitsproduction,accumulation,trafficking,andfunctionareinsufficientlyunderstoodduetothelackofsensitiveandspecificH2O2sensorsthatcanbeusedinlivecells.ThelimitationsofcurrentlyavailableH2O2-responsiveprobesincludeinterferingbackgroundfluorescencefromotherROS,theneedforanexternalactivatingenzyme,lackofwatersolubilityorcompatibility,and/orexcitationprofilesintheultravioletregion.OxiVisionGreen™hydrogenperoxidesensorisnon-fluorescentanddisplaysnoabsorptioninthevisibleregion.TheadditionofH2O2triggersapromptfluorescenceincreasewithconcomitantgrowthofavisiblewavelengthabsorptionband.Thisprobehasalargedynamicrangeowingtoitsbinaryabsorption/emissionresponse.ThefluorescenceresponseofOxiVisionGreen™hydrogenperoxidesensorisH2O2-selective.OxiVisionGreen™hydrogenperoxidesensorexhibitsa>100-foldselectivityforH2O2oversimilarROSsuchasO2-,NO,or-OCl.
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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.PrepareOxiVisionGreenTMhydrogenperoxidesensorworkingsolution:

1.1   Preparea2to5mMstocksolutionofOxiVisionGreenTMhydrogenperoxidesensorinhigh-quality,anhydrousDMSO.Thestocksolutionshouldbeusedpromptly;anyremainingsolutionshouldbealiquotedandfrozenat-20oC.

Note:Avoidrepeatedfreeze-thawcycles.

1.2   Preparea2XOxiVisionGreenTMhydrogenperoxidesensorworkingsolution:Onthedayoftheexperiment,eitherdissolveOxiVisionGreenTMhydrogenperoxidesensorsolidinDMSOorthawanaliquotofthesensorstocksolutiontoroomtemperature.Preparea2Xworkingsolutionattheconcentrationrangingfrom2to20µMin20mMHepesbufferorbufferofyourchoice,pH7.ItisrecommendedtouseOxiVisionGreenTMhydrogenperoxidesensoratthefinalconcentrationof5µMtomeasureH2O2concentrationinsolution.

 

2.RunH2O2Assayinsupernatants:

2.1   Add50µLof2XOxiVisionGreenTMhydrogenperoxidesensorworkingsolution(fromStep1.2)toeachwelloftheH2O2standard,blankcontrol,andtestsamplestomakethetotalH2O2assayvolumeof100 µL/well.

Note:Fora384-wellplate,add25µLofsampleand25µLof2XOxiVisionGreenTMhydrogenperoxidesensorworkingsolutionintoeachwell.

2.2   Incubatethereactionatroomtemperaturefor15to60minutes,protectedfromlight.

2.3   MonitorthefluorescenceincreasewithafluorescenceplatereaderatEx/Em=490/525nm.

2.4   Thefluorescenceinblankwells(withtheassaybufferonly)isusedasacontrol,andissubtractedfromthevaluesforthosewellswiththeH2O2reactions.

 

3.RunH2O2AssayinLiveCells:

OxiVisionGreenTMhydrogenperoxidesensorcanbeloadedpassivelyintolivingcellsandreportthemicromolarchangesinintracellularH2O2concentrations.Thefollowingisasuggestedmicroscopeimagingprotocolwhichcanbemodifiedaccordingtoyourspecificresearchneeds.

3.1   TheOxiVisionGreenTMhydrogenperoxidesensorworkingsolutionshouldbepreparedasStep1.2.ItisrecommendedtousePBSorHanksBalancedSaltSolution(HBSS)with20mMHepesbufferinsteadof20mMHepesbufferonly.

3.2   Treatthecellsasdesired.

3.3   IncubatethecellswithOxiVisionGreenTMhydrogenperoxidesensorworkingsolutionfor5to60minoradesiredperiodoftime.WashthecellswithPBSbuffertwice.

3.4   MonitorthefluorescenceincreaseatEx/Em=490/525nmwithafluorescenceplatereaderwithbottomreadmode.OrimagethefluorescencechangebyafluorescencemicroscopyusingtheFITCchannel.

References&Citations
CitationExplorer

Semaphorin4Dinhibitsneutrophilactivationandisinvolvedinthepathogenesisofneutrophil-mediatedautoimmunevasculitis
Authors:MasayukiNishide,SatoshiNojima,DaisukeIto,HyotaTakamatsu,ShoheiKoyama,SujinKang,TetsuyaKimura,KeikoMorimoto,TakashiHosokawa,YoshitomoHayama
Journal:AnnalsoftheRheumaticDiseases(2017):annrheumdis--2016

Aggravationofbraininfarctionthroughanincreaseinacroleinproductionandadecreaseinglutathionewithaging
Authors:TakeshiUemura,KentaWatanabe,MisakiIshibashi,RyotaroSaiki,KyoshiroKuni,KazuhiroNishimura,ToshihikoToida,KeikoKashiwagi,KazueiIgarashi
Journal:Biochemicalandbiophysicalresearchcommunications(2016):630--635

Hydrogenperoxidedetectionwithhighspecificityinlivingcellsandinflamedtissues
Authors:LeiRong,ChiZhang,QiLei,Ming-MingHu,JunFeng,Hong-BingShu,YiLiu,Xian-ZhengZhang
Journal:RegenerativeBiomaterials(2016):rbw022

Modificationoflignininsugarcanebagassebyamonocopperhydrogenperoxide-generatingoxidasefromThermobifidafusca
Authors:Cheng-YuChen,Cheng-ChengLee,Hung-ShuanChen,Chao-HsunYang,Shu-PingWang,Jyh-HorngWu,MenghsiaoMeng
Journal:ProcessBiochemistry(2016):1486--1495

Dopamine-mediatedoxidationofmethionine127inα-synucleincausescytotoxicityandoligomerizationofα-synuclein
Authors:KazuhiroNakaso,NaokoTajima,SatoruIto,MariTeraoka,AtsushiYamashita,YosukeHorikoshi,DaisukeKikuchi,ShinsukeMochida,KenjiNakashima,TatsuyaMatsura
Journal:PLoSOne(2013):e55068

Hydrogenperoxidestimulatestheepithelialsodiumchannelthroughaphosphatidylinositide3-kinase-dependentpathway
Authors:He-PingMa
Journal:JournalofBiologicalChemistry(2011):32444--32453


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