表观恩特/表观恩特酶胸腺嘧啶DNA糖苷酶（TDG）活性/抑制分析试剂盒（比色法）/P-3094-96/96分析-免疫在线蚂蚁淘旗下平台

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商品详细表观恩特/表观恩特酶胸腺嘧啶DNA糖苷酶（TDG）活性/抑制分析试剂盒（比色法）/P-3094-96/96分析

表观恩特/表观恩特酶胸腺嘧啶DNA糖苷酶（TDG）活性/抑制分析试剂盒（比色法）/P-3094-96/96分析

商品编号： P-3094-96

品牌： Epigentek Group Inc

市场价： ¥11780.00

美元价： 7068.00

产地：美国(厂家直采)

公司：

产品分类：酸碱缓冲液

公司分类： acid_base_buffer_solution

联系Q Q： 3392242852

电话号码： 4000-520-616

电子邮箱： info@ebiomall.com

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商品介绍

InputType:	NuclearExtracts,PurifiedEnzyme
ResearchArea:	DNAMethylation
TargetApplication:	ActivityMeasurement
VesselFormat:	96-WellPlate
100%Guarantee:	6months

ProductOverview

TheEpigenase™ThymineDNAGlycosylase(TDG)Activity/InhibitionAssayKit(Colorimetric)isacompletesetofoptimizedbuffersandreagentsformeasuringtheactivity/inhibitionoftotalTDGenzymeinnuclearextractsorpurifiedTDGenzymesfromabroadrangeofspeciessuchasmammals,plants,fungi,andbacteria,inavarietyofformsincludingbutnotlimitedtoculturedcellsandfreshandfrozentissues.Thekithasthefollowingadvantagesandfeatures:

Colorimetricassaywitheasy-to-followstepsforconvenienceandspeed.Theentireprocedurecanbefinishedwithin5hours.
DirectlymeasuresTDGactivitywithouttheneedforDNAcleavage,electrophoresis,orchromatography.
Bothcell/tissueextractsandpurifiedTDGproteinscanbeused,whichallowsfordetectionofinhibitoryeffectsofTDGinhibitorsinvivoandinvitro.
Novelassayprincipleallowshighsensitivitytobeachieved.Theactivitycanbedetectedfromaslowas20ngofpurifiedTDGprotein.
TDGassaystandardisconvenientlyincluded,whichallowsforspecificquantificationofTDGactivity.
StripmicroplateformatmakestheassayflexIBLe:manualorhighthroughputanalysis(96assays).

BackgroundInformation
DNAdemethylationhasbeendemonstratedtooccurthroughatleasttwodifferentpathways:(1)activedeaminationof5-mCtoTbyanAID/APOBECenzyme,givingaG/TmismatchthatisconvertedtoG/CbythymineDNAglycosylase(TDG)andthensubsequentlybaseexcisionrepair(BER);and(2)iterativeoxidationof5-mCbyTETenzymeto5-hmCandfurtherto5-formycytosine(5-fC)and5-carboxylcytosine(5-caC).Both5-fCand5-caCcanberapidlyexcisedbyTDGtoallowsubsequentBERprocessingwhichresultsin5-fCand5-caCconvertingbacktounmodifiedcytosine.

TDGbelongstotheTDG/mugDNAglycosylasefamily.BesidesplayingacriticalroleinactiveDNAmethylation,TDGalsoremovesthyminemoietiesfromG/Tmismatchesbyhydrolyzingthecarbon-nitrogenbondbetweenthesugar-phosphatebackboneofDNAandthemismatchedthymine.IthasbeenshownthatTDGparticipatesintheregulationofembryonicandgermcelldevelopment,mediatescellulardefenseagainstgeneticandepigeneticmutations,andmediatesDNA-directedcytotoxicity.Inaddition,TDGisaproposedtumorsuppressorcandidate.Thus,themeasurementofTDGactivitywouldbeimportantasTDGmaybeinvolvedinpreventingtumOrigenesisanddeterminingcellularresponsetochemotherapythroughDNAdemethylationandBER,therebybenefitingCancerdiagnosticsandnewtarget-basedcancertherapeutics.

ThecurrentlyusedmethodsforTDGactivitydetectioninvolveDNAcleavagefollowedbyelectrophoresis,whichcanbetimeconsumingandinconvenient,withlowthroughputandhighcost.Toaddressthisissue,EpigentekdevelopedandofferstheEpigenase™ThymineDNAGlycosylase(TDG)Activity/InhibitionAssayKit(Colorimetric).

Principle&Procedure
Inthisassay,a5-fCDNAsubstrateisstablycoatedontomicroplatewells.ActiveTDGbindstothesubstrateandexcises5-fCfromthesubstratebyanickingreaction.Theremainingun-excised5-fCinthesubstratewillberecognizedwithahighaffinity5-fCDNAantibody.Theratiooramountof5-fCinthesubstrate,whichisinverselyproportionaltoenzymeactivity,canthenbecolorimetricallymeasuredbyreADIngtheabsorbanceinamicroplatespectrophotometeratawavelengthof450nm.TheactivityoftheTDGenzymeisinturninverselyproportionaltotheopticaldensityintensitymeasured.

StartingMaterials&InputAmount
Theamountofnuclearextractsforeachassaycanbe2µgto20µgwithanoptimalrangeof5-6µg.Theamountofpurifiedenzymescanbe20ngto1µg,dependingonthepurityandcatalyticactivityoftheenzymes.

Fig.1.SchematicprocedureoftheEpigenase™ThymineDNAGlycosylase(TDG)Activity/InhibitionAssay(Colorimetric).

Fig.2. IllustratedstandardcurvegeneratedwiththeTDGassaystandardfromthe Epigenase™ThymineDNAGlycosylase(TDG)Activity/InhibitionAssay(Colorimetric).

Fig.3. DemonstrationofhighsensitivityandspecificityoftheTDGactivityassayachievedbyusingnuclearextractswiththetheEpigenase™ThymineDNAGlycosylase(TDG) Activity/InhibitionAssayKit(Colorimetric).HelaNuclearextractswereusedintheassay.

ProductComponents

WB(10XWashBuffer)
TAB(TDGAssayBuffer)
BS(BindingSolution)
TS(10XTDGSubstrate)*
TAS(TDGAssayStandard,10µg/ml)*
CA(CaptureAntibody,1000X)*
DA(DetectionAntibody,2000X)*
ES(EnhancerSolution)*
DS(DeveloperSolution)
SS(StopSolution)
8-WellAssayStrips(WithFrame)
UserGuide

*Spinthesolutiondowntothebottompriortouse.

品牌介绍

Epigentek快速，有效地制备DNA，蛋白质和染色质对于进行下游分析至关重要。我们的样品制备套件套件几乎适用于所有工作流程，无论是为ChIP-Seq或ChIP-PCR应用分离染色质，还是从各种起始原料制备亚硫酸氢盐-Seq或MS-PCR的DNA。我们的样品制备套件使您可以直接从提取到反应，为您的实验工作流程提供无数种可能性和灵活性。我们的DNA，蛋白质和染色质制备试剂盒是无数研究机会和下游应用的起点。

公司介绍

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官网：http://www.epigentek.com