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AntibodiesOnline/Oxidative DNA Damage ELISA Kit/ABIN2344961/5 x 96 tests188bio精品生物—专注于实验室精品爆款的电商平台 - 蚂蚁淘旗下精选188款生物医学科研用品
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AntibodiesOnline/Oxidative DNA Damage ELISA Kit/ABIN2344961/5 x 96 tests
Antigen
OxidativeDNADamage
Reactivity
Others
MethodType
CompetitionELISA
Application
ELISA
Options
Bulkdiscount
Supplier
SupplierProductNo.
PurposeTheOxiSelect™OxidativeDNADamageELISAkitisacompetitiveELISAforthequantitativemeasurementof8-OHdG.Theunknown8-OHdGsamplesor8-OHdGstandardsarefirstaddedtoan8-OHdG/BSAconjugatepreabsorbedmicroplate.Afterabriefincubation,ananti-8-OHdGmonoclonalantibodyisadded,followedbyanHRPconjugatedsecondaryantibody.The8-OHdGcontentinunknownsamplesisdeterminedbycomparisonwithpredetermined8-OHdGstandardcurve.
BrandOxiSelect™
SampleTypeUrine,Plasma,CellSamples,Serum,TissueSamples
AnalyticalMethodQuantitative
DetectionMethodColorimetric
Sensitivity100pg/mL8-OHdG
CharacteristicsTheOxiSelect™OxidativeDNADamageELISAKitisacompetitiveenzymeimmunoassaydevelopedforrapiddetectionandquantitationof8-OHdGinurine,serum,orothercellortissueDNAsamples.Thequantityof8-OHdGinunknownsampleisdeterminedbycomparingitsabsorbancewiththatofaknown8-OHdGstandardcurve.Thekithasan8-OHdGdetectionsensitivityrangeof100pg/mLto20 ng/mL.Eachkitprovidessufficientreagentstoperformupto96assays,includingstandardcurveandunknownsamples.
Components
  1. 96-wellProteinBindingPlate:Onestrip-well96wellmicroplate.
  2. Anti-8-OHdGAntibody:One15μLvialofanti-8-OHdG.
  3. SecondaryAntibody,HRPConjugate(1000X):One20μLvial.
  4. AssayDiluent:One50mLbottle.
  5. 10XWashBuffer:One100mLbottle.
  6. SubstrateSolution:One12mLamberbottle.
  7. StopSolution(Part.No.310808):One12mLbottle.
  8. 8-OHdGStandard:One100μLvialof2μg/mL8-OHdGin1XPBS,0.1%BSA.

Box2(shippedonblueicepacks)

Materialnotincluded
  1. 8-OHdGsamplessuchasserum,plasma,urine,orDNAextractedfromcellsortissues
  2. DNAExtractionKit
  3. SodiumAcetate,pH5.2
  4. TrisBuffer,pH7.5
  5. NucleaseP1,AlkalinePhosphatase
  6. 10μLto1000μLadjustablesinglechannelmicropipetteswithdisposabletips
  7. 50μLto300μLadjustablemultichannelmicropipettewithdisposabletips
  8. Multichannelmicropipettereservoir
  9. Microplatereadercapableofreadingat450nm(620nmasoptionalreferencewavelength)
BackgroundFreeradicalsandotherreactivespeciesareconstantlygeneratedinvivoandcauseoxidativedamagetobiomolecules,aprocessheldincheckonlybytheexistenceofmultipleantioxidantandrepairsystemsaswellasthereplacementofdamagednucleicacids,proteinsandlipids.DNAisprobablythemostbiologicallysignificanttargetofoxidativeattack,anditiswidelythoughtthatcontinuousoxidativedamagetoDNAisasignificantcontributortotheage-relateddevelopmentofthemajorcancers,suchasthoseofthecolon,breast,rectum,andprostate.AmongnumeroustypesofoxidativeDNAdamage,theformationof8-hydroxydeoxyguanosine(8-OHdG)isaubiquitousMarkerofoxidativestress.8-OHdG,oneoftheoxidativeDNAdamagebyproducts,isphysiologicallyformedandenhancedbychemicalcarcinogens.DuringtherepairofdamagedDNAinvivobyexonucleases,theresulting8-OH-dGisexcretedwithoutfurthermetabolismintourine.
Comment

  • Detectaslittleas100pg/mLof8-OHdG
  • Suitableforusewithurine,serum,cellsortissues
  • 8-OHdGstandardincludedforabsolutequantitation

PlateUncoated
ReagentPreparation
  • 8-OHdGCoatedPlate:Dilutetheproperamountof8-OHdGConjugate(1 mg/mL)to1 μg/mLin1XPBS.Add100μLofthe1 μg/mL8-OHdGConjugatetoeachwellandincubate3overnightat4 °C.Removethe8-OHdGcoatingsolutionandwashoncewithdH2O.Blotplateonpapertowelstoremoveexcessfluid.Add200μLofAssayDiluenttoeachwellandblockfor1hratroomtemperature.Transfertheplateto4 °CandremovetheAssayDiluentimmediatelybeforeuse.Note:The8-OHdGcoatedwellsarenotstableandshouldbeusedwithin24hrsaftercoating.Onlycoatthenumberofwellstobeusedimmediately.
  • 1XWashBuffer:Dilutethe10XWashBufferto1Xwithdeionizedwater.Stirtohomogeneity.
  • Anti-8-OHdGAntibodyandSecondaryAntibody:ImmediatelybeforeusedilutetheAnti-8-OHdGAntibody1:500andSecondaryAntibody1:1000withAssayDiluent.Donotstoredilutedsolutions.PreparationofStandardCurvePrepareadilutionseriesof8-OHdGstandardsintheconcentrationrangeof0 ng/mLto20 ng/mLbydilutingthe8-OHdGStandardinAssayDiluent(Table1).AssayDiluent8-OHdGStandardTubes8-OHdGStandard(μL)(μL)(ng/mL)110990202500ofTube#1500103500ofTube#250054500ofTube#35002.55500ofTube#45001.256500ofTube#55000.6257500ofTube#65000.3138500ofTube#75000.1569500ofTube#85000.0781005000Table1.Preparationof8-OHdGStandards
SamplePreparation

I.Urine,PlasmaorSerumSamplesClearurine,plasmaorserumsamplescanbedilutedinAssayDiluentanduseddirectlyintheassay.Samplescontainingprecipitatesshouldbecentrifugedat3000gfor10 minutes,orfilteredthrough0.45μmfilter,priortouseintheassay.Note:Formouseorratserumorplasmasamplesitishighlyrecommendedtofilterthesamplewitha10 kDaspinfilterpriortotesting.4II.CellorTissueDNASamples

  1. ExtractDNAfromcellortissuesamplesbyadesiredmethodorcommercialDNAExtractionkit.
  2. DissolveextractedDNAinwaterat1-5 mg/mL.
  3. ConvertDNAsampletosingle-strandedDNAbyincubatingthesampleat95 °Cfor5 minutesandrapidlychillingonice.
  4. DigestDNAsampletonucleosidesbyincubatingthedenaturedDNAwith5-20 unitsofnucleaseP1for2hrsat37 °Cinafinalconcentrationof20mMSodiumAcetate, pH5.2,followedbytreatmentof5-10 unitsofalkalinephosphatasefor1hrat37 °Cinafinalconcentrationof100mMTris, pH7.5.
  5. Thereactionmixtureiscentrifugedfor5 minutesat6000gandthesupernatantisusedforthe8-OHdGELISAassay.

AssayProcedure
  1. Prepareandmixallreagentsthoroughlybeforeuse.Each8-OHdGsampleincludingunknownandstandardshouldbeassayedinduplicate.Highcontent8-OHdGurineorserumsamplesshouldbedilutedatleast10-20foldinAssayDiluent.
  2. Add50μLofunknownsampleor8-OHdGstandardtothewellsofthe8-OHdGConjugatecoatedplate.Incubateatroomtemperaturefor10 minutesonanorbitalshaker.
  3. Add50μLofthedilutedanti-8-OHdGantibodytoeachwell,incubateatroomtemperaturefor1houronanorbitalshaker.
  4. Washmicrowellstrips3timeswith250μL1XWashBufferperwellwiththoroughaspirationbetweeneachwash.Afterthelastwash,emptywellsandtapmicrowellstripsonabsorbentpadorpapertoweltoremoveexcess1XWashBuffer.
  5. Add100μLofthedilutedSecondaryAntibody-EnzymeConjugatetoallwells.
  6. Incubateatroomtemperaturefor1houronanorbitalshaker.
  7. Washmicrowellstrips3timesaccordingtostep4above.Proceedimmediatelytothenextstep.
  8. WarmSubstrateSolutiontoroomtemperature.Add100μLofSubstrateSolutiontoeachwell,includingtheblankwells.Incubateatroomtemperatureonanorbitalshaker.Actualincubationtimemayvaryfrom2-30 minutes.Note:Watchplatecarefully,ifcolorchangesrapidly,thereactionmayneedtobestoppedsoonertopreventsaturation.5
  9. Stoptheenzymereactionbyadding100μLofStopSolutionintoeachwell,includingtheblankwells.Resultsshouldbereadimmediately(colorwillfadeovertime).
  10. Readabsorbanceofeachmicrowellonaspectrophotometerusing450nmastheprimarywavelength.
RestrictionsForResearchUseonly
HandlingAdviceAvoidmultiplefreeze/thawcycles.
Storage-20°C/-80°C
StorageCommentUponreceipt,aliquotandstorethe8-OHdGStandardat-20°Candthe8-OHdGConjugateat-80°Ctoavoidmultiplefreeze/thawcycles.Storeallothercomponentsat4°C.
SupplierImages
ELISA image for Oxidative DNA Damage ELISA Kit (ABIN2344960)8-OHdGELISAStandardCurve
ELISA image for Oxidative DNA Damage ELISA Kit (ABIN2344960)8-OHdGLevelsinHumanUrine.
Productcitedin:Tan,Song,Zhou,Hu:"Antibiotictigecyclineenhancescisplatinactivityagainsthumanhepatocellularcarcinomathroughinducingmitochondrialdysfunctionandoxidativedamage."in:Biochemicalandbiophysicalresearchcommunications,Vol.483,Issue1,pp.17-23,2017(PubMed).

Novaes,Santos,Fialho,Gonçalves,Sequetto,Talvani,Gonçalves:"Nonsteroidalanti-inflammatoryismoreeffectivethananti-oxidanttherapyincounteractingoxidative/nitrosativestressandheartdiseaseinT.cruzi-infectedmice."in:Parasitology,pp.1-13,2017(PubMed).

Chen,Wang,Shen,Lin,Li:"Anthelminthicdrugniclosamidesensitizestheresponsivenessofcervicalcancercellstopaclitaxelviaoxidativestress-mediatedmTORinhibition."in:Biochemicalandbiophysicalresearchcommunications,Vol.484,Issue2,pp.416-421,2017(PubMed).

Wu,Ravikumar,Nguyen,Hsia,Hong:"Lungprotectionbyinhalationofexogenoussolubilizedextracellularmatrix."in:PLoSONE,Vol.12,Issue2,pp.e0171165,2017(PubMed).

AbuBakar,Sarmidi,Tan,MohamadRosdi:"Celastrolattenuatesmitochondrialdysfunctionandinflammationinpalmitate-mediatedinsulinresistanceinC3Ahepatocytes."in:Europeanjournalofpharmacology,Vol.799,pp.73-83,2017(PubMed).

Nelson,Lahiri,Chow,Byrne,Hargett,Breen,Olzomer,Wu,Cooney,Turner,James,Slack-Davis,Lackner,Caldwell,Hoehn:"Inhibitionofhepaticlipogenesisenhanceslivertumorigenesisbyincreasingantioxidantdefenceandpromotingcellsurvival."in:Naturecommunications,Vol.8,pp.14689,2017(PubMed).

Carvalho-Silva,Gomes,Scaini,Rebelo,Damiani,Pereira,Andrade,Gava,Valvassori,Schuck,Ferreira,Streck:"Omega-3fattyacidsupplementationdecreasesDNAdamageinbrainofratssubjectedtoachemicallyinducedchronicmodelofTyrosinemiatypeII."in:Metabolicbraindisease,2017(PubMed).

Kim,Toyono,Berlinicke,Zack,Jurkunas,Usui,Jun:"ScreeningandCharacterizationofDrugsThatProtectCornealEndothelialCellsAgainstUnfoldedProteinResponseandOxidativeStress."in:Investigativeophthalmology&visualscience,Vol.58,Issue2,pp.892-900,2017(PubMed).

Wu,Li,Zhu,Wang,Dai,Zhang,Zheng,Xu,Wang,Zhang,Zhou,Zhang,Shi:"Mdivi-1AlleviatesEarlyBrainInjuryAfterExperimentalSubarachnoidHemorrhageinRats,PossiblyviaInhibitionofDrp1-ActivatedMitochondrialFissionandOxidativeStress."in:Neurochemicalresearch,Vol.42,Issue5,pp.1449-1458,2017(PubMed).

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