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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/Amplite™ Colorimetric Malondialdehyde (MDA) Quantitation Kit/10070/200 Tests
产品编号:10070
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场      地:美国(厂家直采)
联系QQ:1570468124
电话号码:4000-520-616
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美  元  价:$5003.00
品      牌: AAT Bioquest
公      司:AAT Bioquest
公司分类:
AAT Bioquest/Amplite™ Colorimetric Malondialdehyde (MDA) Quantitation Kit/10070/200 Tests
商品介绍
Overview
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Ex/Em(nm)695/None
MWN/A
CAS#N/A
SolventN/A
StorageF/D/L
CategorySmallMoleculeDetection
DiagnosticMolecules
Related
Malondialdehyde(MDA)isnaturalbyproductoflipidperoxidationandiswidelyusedasakeyindicatortodeterminetheoxidativestressandfreerADIcalformation.MeasurementofMDAhashistoricallyreliedonthereactionwiththiobarbituricacid(TBA)toresultsinacompoundthatcanbemeasuredcolorimetricallyat532nmorfluorimetricallyatEx/Em=530nm/550nm.ButthisassayisnotspecifictoMDAandalsotakesplaceunderacidicconditionsat90-100°C.TherehavebeenanumberofcommercialELISAkits,whichmakesitmoreexpensiveandtedious.TheAmplite™ColorimetricMalondialdehyde(MDA)QuantitationKitoffersthemostrapidandconvenientmethodtomeasureMDAwithouttheTBARSheatingsteps.MDABlue™reactswithMDAtogenerateabluecolorproductwhichismeasuredat695nmwithabsorbancemicroplatereader.ThisassayisveryfastandspecifictoMDAwithlittleinterferencefromotheraldehydes.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.
  1. PrepareseriallydilutedMDAstandardsandtestsamples:
    1. Add100uLofddH2OintoMDAStandardvial(ComponentC)tomake100mMMDAstocksolution.
    2. PrepareMDAstandarddilutions:Add4μLof100mMMDAstandard(fromstep1.1)into996μLofDilutionBuffer(ComponentB)toget400μMMDAsolution.Thenperform1:2serialdilutionsindilutionbuffertoget200,100,50,25,12.5,6.25and0μMseriallydilutedMDAstandards.
    3. AddMDAcontainingsamplesandseriallydilutedMDAstandardsintoa96-wellclearbottommicroplateaccordingtoTables1and2.
  2. Table1.LayoutofMDAstandardsandtestsamplesina96-wellclearbottommicroplate
    BLBLTSTS......
    MDA1MDA1............
    MDA2MDA2
    MDA3MDA3
    MDA4MDA4
    MDA5MDA5
    MDA6MDA6
    MDA7MDA7
    Note1:MDA=MDAStandard,BL=BlankControl(dilutionbuffer),TS=TestSample.
    Note2:AddtheserialdilutionsofMDAstandardsfrom6.25μMto400μMintowellsfromMDA1toMDA7.

    Table2.Reagentcompositionforeachwell
    MDAStandardBlankControlTestSample
    SerialDilutions:50µLAssayBuffer:50µL50µL


  3. RunMDAassay:
    1. Add10μLofMDABlue™(ComponentA)solutionintoeachwellofMDAstandard,blankcontrol,andtestsamples(seeStep1.3).
      Note:Fora384-wellplate,add25μLofsample,5μLofMDABlue™(ComponentA)solutionintoeachwell.
      Note:PleasealiquotComponentAintosingleusesizeandstoreunusedat-20°Candavoidlight!
    2. Incubatethereactionmixtureatroomtemperaturefor10-30minutes.
    3. Add40μLofReactionSolution(ComponentD)tomakethetotalassayvolumeof100µL/well.
    4. Incubatethefinalreactionmixtureatroomtemperaturefor30-60minutes.
    5. Monitorabsorbanceincreasewithanabsorbanceplatereaderwithpath-checkcorrectionatODof695~700nm.
References&Citations
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  1. TsikasD.(2016)Assessmentoflipidperoxidationbymeasuringmalondialdehyde(MDA)andrelativesinBIOLOGicalsamples:Analyticalandbiologicalchallenges.AnalBiochem.http://dx.doi.org/10.1016/j.ab.2016.10.021
  2. SteppelerC,HaugenJE,RodbottenR,KirkhusB.(2016)FormationofMalondialdehyde,4-Hydroxynonenal,and4-HydroxyhexenalduringinVitroDigestionofCookedBeef,Pork,Chicken,andSalmon.JAgricFoodChem,64,487.
  3. TsikasD,RothmannS,SchneiderJY,SuchyMT,TrettinA,ModunD,StukeN,MaassenN,FrolichJC.(2016)Development,validationandbiomedicalapplicationsofstable-isotopedilutionGC-MSandGC-MS/MStechniquesforcirculatingmalondialdehyde(MDA)afterpentafluorobenzylbromidederivatization:MDAasabioMarkerofoxidativestressanditsrelationto15(S)-8-iso-prostaglandinF2alphaandnitricoxide(NO).JChromatogrBAnalytTechnolBiomedLifeSci,1019,95.
  4. SpirlandeliAL,DeminiceR,JordaoAA.(2014)Plasmamalondialdehydeasbiomarkeroflipidperoxidation:effectsofacuteexercise.IntJSportsMed,35,14.
  5. MoselhyHF,ReidRG,YousefS,BoyleSP.(2013)Aspecific,accurate,andsensitivemeasureoftotalplasmamalondialdehydebyHPLC.JLipidRes,54,852.
  6. PuntelRL,RoosDH,GrottoD,GarciaSC,NogueiraCW,RochaJB.(2007)AntioxidantpropertiesofKrebscycleintermediatesagainstmalonatepro-oxidantactivityinvitro:acomparativestudyusingthecolorimetricmethodandHPLCanalysistodeterminemalondialdehydeinratbrainhomogenates.LifeSci,81,51.
  7. LykkesfeldtJ.(2007)Malondialdehydeasbiomarkerofoxidativedamagetolipidscausedbysmoking.ClinChimActa,380,50.
  8. GrottoD,SantaMariaLD,BoeiraS,ValentiniJ,CharaoMF,MoroAM,NascimentoPC,PomblumVJ,GarciaSC.(2007)Rapidquantificationofmalondialdehydeinplasmabyhighperformanceliquidchromatography-visIBLedetection.JPharmBiomedAnal,43,619.
  9. FaschingerC,SchmutO,WachswenderC,MossbockG.(2006)[Glaucomaandoxidativestress.Determinationofmalondialdehyde--aproductoflipidperoxidation].Ophthalmologe,103,953.
  10. CelecP,HodosyJ,CelecovaV,VodrazkaJ,CervenkaT,HalcakL,BozekP,KopaniM,KudelaM.(2005)Salivarythiobarbituricacidreactingsubstancesandmalondialdehyde--theirrelationshiptoreportedsmokingandtoparodontalstatusdescribedbythepapillarybleedingindex.DisMarkers,21,133.

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