| InputType: | DNA |
| ResearchArea: | DNAMethylation |
| TargetApplication: | AmountQuantitation |
| VesselFormat: | 96-WellPlate |
| 100%Guarantee: | 6months |
Werecommendusinganewerversionofthisproduct: MethylFlashGlobalDNAMethylation(5-mC)ELISAEasyKit(Colorimetric)
TheMethylFlash™MethylatedDNAQuantificationKit(Colorimetric)isacompletesetofoptimizedbuffersandreagentstocolorimetricallyquantifyglobalDNAmethylationbyspecificallymeasuringlevelsof5-methylcytosine(5-mC)inanELISA-likemicroplate-basedformat.ComparedtoLUMAorLINE-1,Alu,andLTR-basedassays, MethylFlash™ technology directlyquantifiesactualglobalDNAmethylation.
Thiskitisalsospecificallyoptimizedforpairedusewiththe MethylFlashHydroxymethylatedDNAQuantificationKit(Colorimetric) forsimultaneouslyquantifyingbothmethylatedDNAandhydroxymethylatedDNA.
ProductFeatures
TheMethylFlash™MethylatedDNAQuantificationKit(Colorimetric)isafurtherrefinementofourpreviousMethylamp™globalDNAmethylationquantificationkitsby simplifyingtheworkflowandimprovingtheconsistencyofresults.It usesaninnovativemethodtoenablebackgroundsignalstobeextremelylow,eliminatingtheplatedryingandblockingsteps. Comparedtochromatography-basedmethodssuchasHPLCormassspectrometry,thisproductprovidesacost-effectivewaytoaccuratelymeasurelevelsof5-methylcytosine.Thekithasthefollowingadvantagesandfeatures:
- Colorimetricassaywitheasy-to-followstepsforconvenienceandspeed.Theentireprocedurecanbefinishedwithin4hours.
- Innovativekitcompositionenablesbackgroundsignalstobeextremelylow,whicheliminatestheneedforplateblockingandallowstheassaytobesimple,accurate,reliable,andconsistent.
- Highsensitivity,ofwhichthedetectionlimitcanbeaslowas0.2ngofmethylatedDNAand acceptsaslowas50ngofinputgenomicDNA.
- Optimizedantibodyandenhancersolutionsallowhighspecificityto5mC,withnocross-reactivitytounmethylatedcytosineandnoornegligIBLecross-reactivitytohydroxymethylcytosinewithintheindicatedconcentrationrangeofthesampleDNA.
- Universalpositiveandnegativecontrolsareincluded,whicharesuitableforquantifyingmethylatedDNAfromanyspecies.
- Strip-wellmicroplateformatmakestheassayflexible:manualorhighthroughputanalysis.
Principle&Procedure
TheMethylFlash™MethylatedDNAQuantificationKit(Colorimetric)containsallreagentsnecessaryforthequantificationofglobalDNAmethylation.Inthisassay,DNAisboundtostripwellsthatarespecificallytreatedtohaveahighDNAaffinity.ThemethylatedfractionofDNAisdetectedusingcaptureanddetectionantibodiesandthenquantifiedthroughanELISA-likereactionbyreADIngtheabsorbanceinamicroplatespectrophotometerat450nm.TheamountofmethylatedDNAisproportionaltotheODintensitymeasured,whichcanbecalculatedwiththekit"sincludedformulasforrelativemethylationstatusoftwodifferentDNAsamplesorabsolutequantificationof5-methylcytosine(5mC)usingastandardcurve.
Easy,Fast,andFlexible
Theentirecolorimetricassayhaseasy-to-followstepsforconvenienceandspeed,whichcanbecompletedin4hours.Thestrip-wellmicroplateformatallowsforaflexibleassayinmanualorhighthroughputanalysis.UniversalpositiveandnegativecontrolsareincludedwiththekitforquantifyingmethylatedDNAfromanyspeciessuchasmammals,plants,fungi,bacteria,andviruses.
SafeandConvenient
Alltheneededreagents,includingnegativecontrolsandpositivecontrols,forquantificationofglobalDNAmethylationareconvenientlypackagedinthekit.ThedirectcolorimetricquantificationofDNAsamplesreplacesobsoleteorinferiormethodsandeliminatestheneedforDNAdigestion/denaturation,radioactivity,extraction,orchromatography.
HighlySensitiveandSpecific
ThenovelprocedureandproprietarykitcompositionsallowforaccuratequantificationofmethylatedDNAtobeachievedwithhighsensitivityandspecificity.ThedetectionlimitoftheinputDNAcanbeaslowas0.2ngofmethylatedDNA.
ComparisonofAvailableELISA-basedGlobalDNAMethylationAssayMethods
| MethylFlash(Colorimetric) | CompetitorZ | CompetitorC | |
|---|---|---|---|
| AssayPrinciple | DirectELISA | DirectELISA | IndirectELISA |
| Format | 96-wellplate | 96-wellplate | 96-wellplate |
| Sensitivity | Excellentdetectionlimit:0.2ngof5-mCDNA | Gooddetectionlimit:>0.5ngof5-mCDNA | Verypoordetectionlimit:>1000ngof5-mCDNA |
| S/NRatio | >20withverylowbackground | <4withhighbackground | N/A* |
| ProceduralConvenience | NoneedforplateblockinganddenaturationofDNA | RequiresplateblockingandDNAdenaturation | Requiresplateblocking,DNAdigestion,andantibodycompetitiveincubationinanadditionalplate |
| ProtocolTime | <4hr | <4hr | >8hr |
| DNAType&Species | Universalforanyspecies,bothssDNAanddsDNA | HumanandmousessDNAonly | Variousspecies |
| Specificity | Specificto5-mConly | Cross-reactionto5-hmC | N/A* |
| QuantitationType | Bothrelativeandabsolutequantification | Relative | Relative |
| StandardControl | Stable,quantitativeinabsoluteamountof5-mC,andcanbeuniversallyusedforanyspecies | Unstable,cannotbequantitativeinamount,andcanonlybeusedforhumanormouse | N/A* |
| MinimumInputAmount | 50ng | 100ng | 1000ng |
| AccuracyofDetection | High,stableandquantifiedstandardcontrol.ProvenclosecorrelationwithLC-MSanalysisbyusers | Unclear,unstableandun-quantifiedstandardcontrol | N/A* |
| %5-mCcalculation | Simple | Complicated | NotProvided |
| PatentedMethod | Yes | No | No |
| Popularity | Veryhighpublishedcitationcount | Lowpublishedcitationcount | Lowpublishedcitationcount |
| SupportExpertise | Since2006 | Since2013 | Since2011 |
*Insufficientinformationprovidedbydatasheetorproductfailedtoproducedatatomakeconclusivedetermination
ME1(10XWashBuffer)
ME2(BindingSolution)
ME3(NegativeControl,20µg/ml)*
ME4(PositiveControl,20µg/ml)*
ME5(CaptureAntibody,1000µg/ml*
ME6(DetectionAntibody,400µg/ml)*
ME7(EnhancerSolution)*
ME8(DeveloperSolution)
ME9(StopSolution)
8-WellAssayStrips(WithFrame)
UserGuide
*Spinthesolutiondowntothebottompriortouse.
Note:TheME3NegativeControlisanunmethylatedpolynucleotidecontaining50%ofcytosine.TheME4PositiveControlisamethylatedpolynucleotidecontaining50%of5-methylcytosine.
