
Overview | ![]() PrinterFriendlyVersion |
Ex/Em(nm) | 695/None |
MW | N/A |
CAS# | N/A |
Solvent | N/A |
Storage | F/D/L |
Category | SmallMoleculeDetection DiagnosticMolecules |
Related |
- PrepareseriallydilutedMDAstandardsandtestsamples:
- Add100uLofddH2OintoMDAStandardvial(ComponentC)tomake100mMMDAstocksolution.
- PrepareMDAstandarddilutions:Add4μLof100mMMDAstandard(fromstep1.1)into996μLofDilutionBuffer(ComponentB)toget400μMMDAsolution.Thenperform1:2serialdilutionsindilutionbuffertoget200,100,50,25,12.5,6.25and0μMseriallydilutedMDAstandards.
- AddMDAcontainingsamplesandseriallydilutedMDAstandardsintoa96-wellclearbottommicroplateaccordingtoTables1and2.
Table1.LayoutofMDAstandardsandtestsamplesina96-wellclearbottommicroplate - RunMDAassay:
- Add10μLofMDABlue™(ComponentA)solutionintoeachwellofMDAstandard,blankcontrol,andtestsamples(seeStep1.3).
Note:Fora384-wellplate,add25μLofsample,5μLofMDABlue™(ComponentA)solutionintoeachwell.
Note:PleasealiquotComponentAintosingleusesizeandstoreunusedat-20°Candavoidlight! - Incubatethereactionmixtureatroomtemperaturefor10-30minutes.
- Add40μLofReactionSolution(ComponentD)tomakethetotalassayvolumeof100µL/well.
- Incubatethefinalreactionmixtureatroomtemperaturefor30-60minutes.
- Monitorabsorbanceincreasewithanabsorbanceplatereaderwithpath-checkcorrectionatODof695~700nm.
- Add10μLofMDABlue™(ComponentA)solutionintoeachwellofMDAstandard,blankcontrol,andtestsamples(seeStep1.3).
BL | BL | TS | TS | ... | ... | ||||||
MDA1 | MDA1 | ... | ... | ... | ... | ||||||
MDA2 | MDA2 | ||||||||||
MDA3 | MDA3 | ||||||||||
MDA4 | MDA4 | ||||||||||
MDA5 | MDA5 | ||||||||||
MDA6 | MDA6 | ||||||||||
MDA7 | MDA7 |
Note2:AddtheserialdilutionsofMDAstandardsfrom6.25μMto400μMintowellsfromMDA1toMDA7.
Table2.Reagentcompositionforeachwell
MDAStandard | BlankControl | TestSample |
SerialDilutions:50µL | AssayBuffer:50µL | 50µL |
References&Citations | ![]() PrinterFriendlyVersion |
- TsikasD.(2016)Assessmentoflipidperoxidationbymeasuringmalondialdehyde(MDA)andrelativesinBIOLOGicalsamples:Analyticalandbiologicalchallenges.AnalBiochem.http://dx.doi.org/10.1016/j.ab.2016.10.021
- SteppelerC,HaugenJE,RodbottenR,KirkhusB.(2016)FormationofMalondialdehyde,4-Hydroxynonenal,and4-HydroxyhexenalduringinVitroDigestionofCookedBeef,Pork,Chicken,andSalmon.JAgricFoodChem,64,487.
- 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.
- SpirlandeliAL,DeminiceR,JordaoAA.(2014)Plasmamalondialdehydeasbiomarkeroflipidperoxidation:effectsofacuteexercise.IntJSportsMed,35,14.
- MoselhyHF,ReidRG,YousefS,BoyleSP.(2013)Aspecific,accurate,andsensitivemeasureoftotalplasmamalondialdehydebyHPLC.JLipidRes,54,852.
- PuntelRL,RoosDH,GrottoD,GarciaSC,NogueiraCW,RochaJB.(2007)AntioxidantpropertiesofKrebscycleintermediatesagainstmalonatepro-oxidantactivityinvitro:acomparativestudyusingthecolorimetricmethodandHPLCanalysistodeterminemalondialdehydeinratbrainhomogenates.LifeSci,81,51.
- LykkesfeldtJ.(2007)Malondialdehydeasbiomarkerofoxidativedamagetolipidscausedbysmoking.ClinChimActa,380,50.
- GrottoD,SantaMariaLD,BoeiraS,ValentiniJ,CharaoMF,MoroAM,NascimentoPC,PomblumVJ,GarciaSC.(2007)Rapidquantificationofmalondialdehydeinplasmabyhighperformanceliquidchromatography-visIBLedetection.JPharmBiomedAnal,43,619.
- FaschingerC,SchmutO,WachswenderC,MossbockG.(2006)[Glaucomaandoxidativestress.Determinationofmalondialdehyde--aproductoflipidperoxidation].Ophthalmologe,103,953.
- CelecP,HodosyJ,CelecovaV,VodrazkaJ,CervenkaT,HalcakL,BozekP,KopaniM,KudelaM.(2005)Salivarythiobarbituricacidreactingsubstancesandmalondialdehyde--theirrelationshiptoreportedsmokingandtoparodontalstatusdescribedbythepapillarybleedingindex.DisMarkers,21,133.
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1.模板提取(一般为RNA):Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定:分光光度计或NanoDrop
3.逆转录:逆转录试剂盒(或者一步法试剂盒),这一步可以用普通PCR做,也可以用水域做。
4.荧光定量PCR试剂:通常有用SYBR Green Mix做的,但是这里建议你用EvaGreen做,灵敏度和平行性都要好于SYBR Green,并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他:除了以上的那些还需要离心管、PCR管或板(Axygen反应比较好)、移液枪等,暂时就想到这么多。
1.模板提取(一般为RNA):Trizol、氯仿、异丙醇、无水乙醇、DEPC处理水
2.模板浓度测定:分光光度计或NanoDrop
3.逆转录:逆转录试剂盒(或者一步法试剂盒),这一步可以用普通PCR做,也可以用水域做。
4.荧光定量PCR试剂:通常有用SYBR Green Mix做的,但是这里建议你用EvaGreen做,灵敏度和平行性都要好于SYBR Green,并且如果你那是ABI或者Stratagene的PCR如果用SYBR Green还需要加一步Rox很麻烦。
5.其他:除了以上的那些还需要离心管、PCR管或板(Axygen反应比较好)、移液枪等,暂时就想到这么多。
MicroBCA试剂测定范围是0.5-20μg/ml
楼主自己看看经销商价格
临床实验室对商品定量试剂盒分析性能的验证.pdf(17431.6k)
如果要是直接注射,就不知道了

