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当前位置: 首页 > 产品中心 > Other_kits > AAT Bioquest/Amplite™ Red/11011/1000 Assays
商品详细AAT Bioquest/Amplite™ Red/11011/1000 Assays
AAT Bioquest/Amplite™ Red/11011/1000 Assays
AAT Bioquest/Amplite™ Red/11011/1000 Assays
商品编号: 11011
品牌: aatbio
市场价: ¥27560.00
美元价: 16536.00
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
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Ex/Em(nm)571/585
MWN/A
CAS#N/A
SolventDMSO
StorageF/D/L
CategoryEnzymeDetection
HorserADIshPeroxidase(HRP)
RelatedBiochemicalAssays
OurAmplite™Redisasensitivefluorogenicperoxidasesubstratethatgeneratesahighlyredfluorescentproductthathasmaximumabsorptionof571nmandmaximumemissionof585nm.UnlikeotherHRPsubstratessuchasdihydrofluoresceinsanddihydrorhodamines,theair-oxidationofAmplite™Redisminimal.Amplite™RedisoneofthemostsensitiveandstablefluorogenicprobesfordetectingHRPandH2O2.Amplite™RedhasbeenwidelyusedtodetectHRPinmanyimmunoassays.Ontheotherhand,Amplite™RedcanalsobeusedtodetecttraceamountofH2O2.TheAmplite™Red-basedH2O2detectionisatleastoneorderofmagnitudemoresensitivethanthecommonlyusedscopoletinassayforH2O2.BecauseH2O2isproducedinmanyenzymaticredoxreactions,Amplite™Redcanbeusedincoupledenzymaticreactionstodetecttheactivityofmanyoxidasesand/orrelatedenzymes/substratesorcofactorssuchasglucose,acetylcholineandcholesterol,L-glutamate,aminoacids,etc.
SpectrumAdvancedSpectrumViewer

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Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

AssayProtocolforPeroxidase(HRP)withAmplite™Red(forone96wellblackplate)


A1.PrepareAmplite™Redperoxidaseworkingsolution:

1.1   Prepare250XAmplite™Redstocksolution: Add200µLofanhydrousDMSOintothevial,mixedwell.Thestocksolutionshouldbeusedpromptly.Anyunusedsolutionneedtobealiquotedandrefrozenat<-20oC.

Note:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.

1.2   Prepare1XAmpliteTMRedperoxidaseworkingsolution:Onthedayoftheexperiment,eitherdissolveAmpliteTMRedsolidinDMSOorthawanaliquotoftheAmpliteTMRedstocksolutionatroomtemperature.Prepare1Xworkingsolutionbyadding20µLof250XAmpliteTMRedstocksolution(fromStep1.1)in5mLof50mMphosphatebufferorbufferofyourchoice,pH7with200µMH2O2.

Note:AmpliteTMRedisunstableinthepresenceofthiolssuchasDTTandb-mercaptoethanol.Thiolshigherthan10µM(finalconcentration)couldsignificantlydecreasetheassaydynamicrange.NADHandglutathione(reducedfrom:GSH)mayinterferewiththeassay.

 

A2.Runperoxidaseassayinsupernatants:

2.1   Add50µLof1XAmpliteTMRedperoxidaseworkingsolution(fromStep1.2)intoeachwelloftheperoxidase standard,blankcontrol,andtestsamplestomakethetotalperoxidaseassayvolumeof100µL/well.

Note:Fora384-wellplate,add25µLofsampleand25µLof1XAmpliteTMRedperoxidaseworkingsolutionintoeachwell.

2.2   Incubatethereactionatroomtemperaturefor10to30minutes,protectedfromlight.

2.3   MonitorthefluorescenceincreaseatEx/Em=540/590nmwithafluorescenceplatereader.

2.4   Thefluorescenceinblankwells(withtheassaybufferonly)isusedasacontrol,andissubtractedfromthevaluesforthosewellswiththeperoxidasereactions.

AssayProtocolforH2O2withAmplite™Red(forone96wellblackplate)


B1.PrepareAmpliteTMRedH2O2workingsolution:

1.1   Prepare250XAmplite™Redstocksolution:Add200µLofanhydrousDMSOintothevial,mixedwell.Thestocksolutionshouldbeusedpromptly.Anyunusedsolutionneedtobealiquotedandrefrozenat<-20oC.

Note:Avoidrepeatedfreeze-thawcycles,andprotectfromlight.

1.2   Prepare1XAmpliteTMRedH2O2workingsolution:Onthedayoftheexperiment,eitherdissolveAmpliteTMRedsolidinDMSOorthawanaliquotoftheAmpliteTMRedstocksolutionatroomtemperature.Prepare1Xworkingsolutionbyadding20µLof250Xstocksolution(fromStep1)in5mLof50mMphosphatebufferorbufferofyourchoice,pH7with0.4units/mLperoxidase.

Note:AmpliteTMRedisunstableinthepresenceofthiolssuchasDTTandb-mercaptoethanol.Thiolshigherthan10µM(finalconcentration)couldsignificantlydecreasetheassaydynamicrange.NADHandglutathione(reducedfrom:GSH)mayinterferewiththeassay.

B2.RunH2O2assayinsupernatants:

2.1   Add50µLof1XAmpliteTMRedH2O2workingsolution(fromStep1.2)intoeachwelloftheH2O2standard,blankcontrol,andtestsamplestomakethetotalH2O2assayvolumeof100µL/well.

Note:Fora384-wellplate,add25µLofsampleand25µLof1XAmpliteTMRedH2O2workingsolutionintoeachwell.

2.2   Incubatethereactionatroomtemperaturefor10to30minutes,protectedfromlight.

2.3   MonitorthefluorescenceincreaseatEx/Em=540/590nmwithafluorescenceplatereader.

2.4   Thefluorescenceinblankwells(withtheassaybufferonly)isusedasacontrol,andissubtractedfromthevaluesforthosewellswiththeH2O2reactions

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


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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