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蚂蚁淘在线 / 品牌中心 / Epigentek / Epigentek/MethylFlash Urine 5-Methylcytosine (5-mC) Quantification Kit (Colorimetric)/P-1039-48/48 assays

Epigentek/MethylFlash Urine 5-Methylcytosine (5-mC) Quantification Kit (Colorimetric)/P-1039-48/48 assays

价格
￥5880.00
货号：P-1039-48
浏览量：127
品牌：Epigentek
服务
全国联保
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商品描述
InputType:Urine
ResearchArea:DNAMethylation
TargetApplication:AmountQuantitation
VesselFormat:96-WellPlate
100%Guarantee:6months
ProductOverview

TheMethylFlash™Urine5-Methylcytosine(5-mC)QuantificationKit (Colorimetric)  isacompletesetofoptimizedbuffersandreagentstocolorimetricallyquantify5-mCinurineusinganinhibitorycompetitiveimmunoassaymethod.It issuitablefordetectingtotalurinary5-methylcytosinelevels,resultingfromwholebodyturnoverordegradationofmethylatedDNA/RNA,inurinefromhumansandanimals.Theurinesamplescanbeinfreshorfrozenform. Thekithasthefollowingadvantagesandfeatures:

  • Innovativecolorimetricassaywitheasy-to-followstepsforconvenienceandspeed.Theentireprocedurecanbefinishedwithin4hours.
  • 96strip-wellmicroplateformatmakestheassayflexIBLe:manualorhighthroughputanalysis.
  • Innovativekitcompositionenablesbackgroundsignalstobeextremelylow,whicheliminatestheneedforplateblockingandallowstheassaytobesimple,accurate,reliable,andconsistent.
  • Thelevelof5-mCmeasuredinhumanurinesamplesusingthiskitiscomparabletothatdetectedbyHPLCmethod. 
  • Anovelassayprincipleallowshighsensitivitytobeachieved.Thedetectionlimitcanbeaslowas0.03ng/assaywellor1nMof5-mC.
  • Lowinputrangeofurineforeachassaywithavolumeof0.5to5µlandanoptimalvolumeof1µl.
  • Optimizedantibodyandenhancersolutionsallowforhighspecificityto5-mC,withoutcross-reactivitytounmethylatedcytosine. 
  • Negativecontrolandpositivestandardareincluded,whicharesuitableforquantificationof5-mCinfreeformand5-mCcontainedinmethylatedDNA/RNAfragmentsfromdifferenturinesamples.

BackgroundInformation
Nucleobase5-methylcytosine(5-mC),amodifiedformofcytosineconvertedbycytosinemethyltransferases,iswidespreadbothinDNAanddifferentcellularRNAs.TheBIOLOGicalimportanceofDNA5-mCmethylationasamajorepigeneticmodificationinphenotypeandgeneexpressionhasbeenwidelyrecognized.RecentdatastronglysuggeststhatRNA5-mCmethylationisalsoinvolvedintheregulationofvariousbiologicalprocessesincludingtRNAstABIlity/recognitionandmRNAtranslation. 

Urinaryexcretionof5-mCincludingboth5-methyl-2-deoxycytidineand5-methylcytidineisanindicationofawholebodyturnoverordegradationofmethylatedDNAandRNA.Theurinary5-mClevelcanbealteredbyachangeofthebodies’turnoverofmethylatedDNA/RNAoralterationofcellularDNA/RNAmethylationstatus.Anumberofstudieshaveindicatedthat5-mCexcretedinurinehasthepotentialtoactasacancerbioMarker,withanincreasedlevelarisingfromdiseaseonsetandprogression.Forexample,anelevatedlevelofurinary5-mCwasobservedinlungcancer,breastcancer,andleukemiapatientswithactivediseasestates.Itwasalsoshownthaturinary5-mCexcretioniseffectedbyAlzheimer’sdiseaseorbyrADIationtreatment.Itissuggestedthaturinary5-mCmightbeapplicableasabiologicalmarkerfordetectingsometypesofcancerandmonitoringcancerprogressionaftertreatmentwithradiationordemethylationreagents. 

Chromatography-basedtechniquessuchasHPLCandTLCmassspectrometryarecommonlyusedfordetecting5-mCinurineandalthoughfairlyacurate,aretimeconsuming,lesssensitive,andhavelowthroughputwithhighcosts.Toaddressthisproblem,EpigentekofferstheMethylFlash™ Urine5-Methylcytosine(5-mC)QuantificationKit (Colorimetric) toquantify5-mCorbodyturnoverstatusofmethylatedDNA/RNAusingurinesamples.

Principle&Procedure
InthisELISA-likeinhibitorycompetitiveimmunoassayassay,urinesamplesandthe5-mCstandardarefirstincubatedwitha5-mCantibodysolutionandthentransferredtostripwellscoatedwithmethylatedDNA(5-mC).Thewelliswashedtoremoveanyunboundreagentsafterincubationandthenadetectionantibodyisaddedtogenerateasignalthatcanbemeasuredcolorimetricallybyreadingtheabsorbanceinamicroplatespectrophotometer.Because5-mCintheurinesampleinhibitsthebindingof5-mCantibodyto5-mCcoatedonthewell,higherconcentrationsof5-mCintheurinesampleleadtoareducedbindingoftheantibodytothe5-mConthewell.ThereforethesignalorODintensitymeasuredfromthewellwillbeinverselyproportionaltotheamountof5-mCintheurinesampleandtheamountof5-mCintheurinesamplecanbequantifiedbyacomparisonwithapredetermined5-mCstandard.


Fig.1.SchematicprocedurefortheMethylFlash™Urine5-MethylcytosineQuantificationKit(Colorimetric).


Fig.2. 5-mCstandardwasaddedintotheassaywellsatdifferentconcentrationsandthenmeasuredwiththeMethylFlash™Urine5-MethylcytosineQuantificationKit (Colorimetric).


Fig.3. 5-mClevelisquantifiedfromdifferentvolumesofhumanurineusingtheMethylFlash™Urine5-MethylcytosineQuantificationKit (Colorimetric).

 

 

ProductComponents

WB(10XWashBuffer)
BS(BindingSolution)
MS(5-mCStandard,4µg/ml)*
ME3(NegativeControl,50µg/ml)*
ME4(5-mCDNAsolution,320X)*
ME5(CaptureAntibody,1000X)*
ME6(DetectionAntibody,2000X)*
ES(EnhancerSolution)*
DS(DeveloperSolution)
SS(StopSolution)
Plate1(AssayPlate)(WithFrame)
Plate2(SamplePreparationPlate)
UserGuide

*Spinthesolutiondowntothebottompriortouse.

 

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