Overview:
| ProductName | DNADamage(8-OHdG)ELISAkit |
| Description | Colorimetricdetectionof8-hydroxy-2-deoxyGuanosine |
| SpeciesReactivity | SpeciesIndependent |
| Platform | Microplate |
| SampleTypes | Celllysates,Plasma,Samplematrices,Urine |
| DetectionMethod | ColorimetricAssay |
| AssayType | CompetitiveELISA(Enzyme-linkedImmunosorbentAssay) |
| Utility | ELISAKitfor8-OHdGdetectioninsamples. |
| Sensitivity | 0.59ng/mL |
| AssayRange | 0.94-60ng/mL |
| Precision | Intra-AssayPrecision:Threesamplesofknownconcentrationwereassayedthirtytimesononeplate;theintra-assaycoefficientofvariationoftheDNADamageELISAhasbeendeterminedtobe<5%. Inter-Assay Precision: Three samples of known concentration were assayed thirty times in three individual assays; the inter-assay coefficient of variation of the DNA Damage ELISA has been determined to be <5%. |
| IncubationTime | 1hour |
| NumberofSamples | 39samplesinduplicate |
| OtherResources | KitBooklet,MSDS,CalculationsWorksheet |
Properties
| StorageTemperature | 4ºCand-20ºC | ||||||||||||||||||||||||||||||
| ShippingTemperature | BlueIce | ||||||||||||||||||||||||||||||
| ProductType | ELISAKits | ||||||||||||||||||||||||||||||
| AssayOverview | 1.PreparestandardandsamplesintheSampleandStandardDiluent.2.Add50µLofpreparedstandardsandsamplesintriplicatetoappropriatewells.3.Add50µLofthedilutedantibodypreparationtotheappropriatewells.4.CoverplatewithPlateCoverandincubateatroomtemperature(20-25°C)for1hour.5.Washplate4timeswith1XWashBuffer.6.Add100µLofTMBSubstratetoeachwell.7.Coverplateanddeveloptheplateinthedarkatroomtemperaturefor30minutes.8.Add100µLofStopSolutiontoeachwell.9.Measureabsorbanceonaplatereaderat450nm.10.Plotthestandardcurveandcalculatesampleconcentrations. | ||||||||||||||||||||||||||||||
| KitOverview |
| ||||||||||||||||||||||||||||||
| CiteThisProduct | DNADamage(8-OHdG)ELISAKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-120) |
BIOLOGicalDescription
| AlternativeNames | 8-OH-dGELISAKit,8OHGELISAKit,80GELISAKit,8hydroxyguanineELISAKit,8-OHdGELISAKit,DNADamageELISAKit |
| ResearchAreas | Cancer,CellSignaling,Oxidation,OxidativeStress,Post-translationalModifications |
| ScientificBackground | 8-hydroxy-2-deoxyGuanosine(8-OH-dG)isproducedbytheoxidativedamageofDNAbyreactiveoxygenandnitrogenspeciesandservesasanestablishedMarkerofoxidativestress(1-4).Hydroxylationofguanosineoccursinresponsetobothnormalmetabolicprocessesandavarietyofenvironmentalfactors(i.e.,anythingthatincreasesreactiveoxygenandnitrogenspecies).Increasedlevelsof8-OH-dGareassociatedwiththeagingprocessaswellaswithanumberofpathologicalconditionsincludingcancer,diabetes,andhypertension(5-9).Incomplexsamplessuchasplasma,celllysates,andtissues,8-OH-dGcanexistaseitherthefreenucleosideorincorporatedinDNA.Oncethebloodentersthekidney,free8-OH-dGisreADIlyfilteredintotheurine,whilelargerDNAfragmentsremaininthebloodstream.Becauseofthecomplexityofplasmasamples,urineisamoresuitablematrixforthemeasurementoffree8-OH-dGthanplasma.Urinarylevelsof8-OH-dGrangebetween2.7-13ng/mgcreatine,whileplasmalevelsoffree8-OH-dGhavebeenreportedtobebetween4-21pg/mlasdeterminedbyLC-MS(10-11). |
| References | 1.MaxeyK.M.,MaddipatiK.R.,BirkmeierJ.(1992)JClinImmunoassay15:116-120. 2.PradellesP.,GrassiJ.,MacloufJ.(1990)MethodsEnzymol.187:24-34. 3.MacloufJ.,GrassiJ.,PradellesP.(1987)DevImmunoassayTechMeaseicosanoids. 4.LinH.,etal.(2004)BiochemJ.380:541-548. 5.BogdanovM.B.,etal.(1999)FreeRadicBiolMed.27(5/6):647-666. 6.LeeJ.,etal.(2005)Hypertension45:986-990. 7.Leinonen,J.,etal.(1997)FEBSLett.417:150-152. 8.EndoK.,etal.(2006)J.Atheroscler.Thromb.13:68-75. 9.KuoH.,etal.(2007)MutatRes.631:62-68. 10.ShenJ.,etal.(2007)Cancer109:574-580. 11.BeckmanK.B.,AmesB.N.(1997)JBiolChem272:19633-19636. 12.EpeB.,etal.(1996)NucleicAcidsRes24:4105-4110. 13.SpencerJ.P.E.,etal.(1995)FEBSLett374:233-236. 14.FloydR.A.(1990)FASEBJ4:2587-2597. |
ProductImages
TypicalStandardCurvefortheDNADamage(8-OHdG)ELISAkit(Enzyme-LinkedImmunosorbentAssay)StressXpress®–SKT-120.AssayType:CompetitiveELISA.DetectionMethod:ColorimetricAssay.AssayRange:0.94–60ng/ml.
ChemicalEquationoftheOxidationofGuanosine
Diagramofthe8-OHdGCompetitiveELISA
UrineSpikeAssay
DiagramofthePreparationofthe8-OHdGStandards
DiagramoftheTriplicateSamplePlateFormat
PreviewoftheCalculationsWorksheet
ProductCitations(43)
OtherCitations
AssociationofMetabolicSyndromeandOxidativeDNADamageinHIV/AIDSPatients.
Kolgiri,V.,Nagar,V.andPatil,V.(2017)IndJClinBiochem.[Epubaheadofprint].
PubMedID:28535982ReactivityHumanApplications:Plasma
AntiretroviralTherapy‑inducedInsulinResistanceandOxidativeDeoxyNucleicAcidDamageinHumanImmunodeficiencyVirus‑1Patients.
VKHonnapurmath,V.W.P.(2017)IndianJEndocrMetab.21:316-21.
PubMedID:N/AReactivityHumanApplications:Blood
Urinaryexcretionofuricacid,allantoin,and8-OH-Deoxyguanosineinuricase-knockoutmice.
Inazawa,K.etal.(2016)NucleosidesNucleotidesNucleicAcids.35(10-12):559-565.
PubMedID:27906613ReactivityMouseApplications:Urine
Proteincarbonylcontent:anovelbiomarkerforaginginHIV/AIDSpatients.
Kolgiri,V.andPatil,V.W.(2016)BrazJInfectDis.pii:S1413-8670(16)30432-9.
PubMedID:27821249ReactivityHumanApplications:Plasma
Coagulin-lamelioratesTLR4inducedoxidativedamageandimmuneresponsebyregulatingmitochondriaandNOX-derivedROS.
Reddy,SS(2016)ToxicolApplPharmacol.309:87-100.
PubMedID:27568862ReactivityMouseApplications:Liverhomogenates
PreventiveEffectofResveratrolagainstBrainMitochondriaDNADamage,LipidPeroxidation,InflammationandSeizuresInducedbyKainicAcidinMice.
Hussein,S.A.,ABDel-mageid,A.D.,Abd-Elhamed,O.M.,Amin,A.andAlharthy,H.S.(2016)IntJPharSci.6(4):1634-1646
PubMedID:N/AReactivityMouseApplications:Braintissue
NeuroprotectiveEffectofCurcuminonKainicAcidModelofEpilepsyinMaleSwissAlbinoMice.
Hussein,S.A.,Abdel-mageid,A.D.,Abd-Elhamed,O.M.,Amin,A.andAlharthy,H.S.(2016)IntJChemNatSci.4(4):447-460.
PubMedID:N/AReactivityMouseApplications:Braintissue
DNAdamageandtheactivationofthep53pathwaymediatealterationsinmetabolicandsecretoryfunctionsofadipocytes.
Vergoni,B.etal.(2016)Diabetes.
PubMedID:27388216ReactivityHumanApplications:Adipocytes
Discoveringbiomarkersforantidepressantresponse:protocolfromtheCanadianbiomarkerintegrationnetworkindepression(CAN-BIND)andclinicalcharacteristicsofthefirstpatientcohort.
Lam,R.W.etal.(2016)BMCPsychiatry.16(1):105.
PubMedID:27084692ReactivityHumanApplications:BlootandUrine
Podocyte-specificNox4deletionaffordsrenoprotectioninamousemodelofdiabeticnephropathy.
Jha,J.C.etal.(2015)Diabetologia.59(2):379-89.
PubMedID:26508318ReactivityMouseApplications:Urine
DoeschronicraiseofmetalionlevelsinduceoxidativeDNAdamageandhypoxia-likeresponseinpatientswithmetal-on-metalhipresurfacing?
Savarino,L.etal.(2015)JBiomedMaterResBApplBiomater.105(2):460-466.
PubMedID:26477446ReactivityHumanApplications:Serum
HigherurinaryLevelsof8-hydroxy-2"-deoxyguanosineareassociatedwithaworseRANKL/OPGratioinpostmenopausalwomenwithosteopenia.
Cervellati,C.etal.(2015)OxidMedCellLong.2016:6038798.
PubMedID:26635910ReactivityHumanApplications:Urine
TurmeroneenrichedstandardizedCurcumalongaextractalleviatesLPSinducedinflammationandcytokineproductionbyregulatingTLR4–IRAK1–ROS–MAPK–NFκBaxis.
Rana,M.etal.(2015)JFunctionalFoods.16(2015):152–163.
PubMedID:N/AReactivityMouseApplications:Liverhomogenates
Synthesis,characterizationandbiologicalactivityofsomeunsymmetricalSchiffbasetransitionmetalcomplexes.
Esmadi,F.T.etal.(2015)DrugChemToxicol.:1-7.
PubMedID:25791998ReactivityHumanApplications:Lymphocytes
Late-interventionstudywithebseleninanexperimentalmodeloftype1diabeticnephropathy.
Tan,S.M.,Sharma,A.,Stefanovic,N.anddeHaan,J.B.(2015)FreeRadicRes.49(3):219-27.
PubMedID:25465090ReactivityMouseApplications:Urine
Immediateanddelayedeffectsofgrowthconditionsonageingparametersinnestlingzebrafinches.
Reichert,S.etal.(2015)JExpBiol.218(Pt3):491-9.
PubMedID:25524985ReactivityZebrafinchApplications:Bloodcells
AderivativeofBardoxolonemethyl,dh404,inaninversedose-dependentmanner,lessensdiabetes-associatedatherosclerosisandimprovesdiabetickidneydisease.
Tan,S.M.,etal.(2014)Diabetes.63(9):3091-103.
PubMedID:24740568ReactivityMiceApplications:Urine
Elevationimpactsthebalancebetweengrowthandoxidativestressincoaltits.
Stier,A.,etal.(2014)Oecologia.175(3):791-800.
PubMedID:24805201ReactivityBird(coaltits)Applications:Redbloodcells
EvaluationofvitaminB12effectsonDNAdamageinducedbypaclitaxel.
Alzoubi,K.,KhabourO.,KhaderM.,MhaidatN.andAl-Azzam,S.(2014)DrugChemToxicol.37(3):276-80.
PubMedID:24215581ReactivityHumanApplications:Blood
EffectofTherapeuticHypothermiaonDNADamageandNeurodevelopmentalOutcomeAmongTermNeonateswithPerinatalAsphyxia:ARandomizedControlledTrial.
Gane,B.D.,etal.(2014)JTropPediatr.60(2):134-40.
PubMedID:24343823ReactivityHumanApplications:Blood
Disruptionofpro-oxidantandantioxidantsystemswithelevatedexpressionoftheubiquitinproteosomesysteminthecachecticheartmuscleofnudemice.
Hinch,E.C.A.,Sullivan-Gunn,M.J.,VaughanV.C.,McGlynnM.A.,Lewandowski,PaulA.(2013)JCachexiaSarcopeniaMuscle.4(4):287-93.
PubMedID:24030522ReactivityMouseApplications:Tumortissue
OxidativeDamageofDNAConfersResistancetoCytosolicNucleaseTREX1DegradationandPotentiatesSTING-DependentImmuneSensing.
Gehrke,N.etal.(2013)Immunity.39(3):482-95.
PubMedID:23993650ReactivityMouseApplications:RMAcells(T-celltumorcells)
Differentialeffectsofdocosahexanoicacid(DHA)onpretermandtermplacentalpro-oxidant/anti-oxidantbalance.
Stark,M.etal.(2013)Reproduction.146(3):243-51.
PubMedID:23813449ReactivityHumanApplications:Placenta
TheModifiedSelenenylAmide,M-hydroxyEbselen,AttenuatesDiabeticNephropathyandDiabetes-AssociatedAtherosclerosisinApoE/GPx1DoubleKnockoutMice.
Tan,S.M.,etal.(2013)PLoSONE.8(7):e69193.
PubMedID:23874911ReactivityMouseApplications:Urine(diabeticandnon-diabetic)
TiO2NanoparticleExposureandIlluminationduringZebrafishDevelopment:MortalityatPartsperBillionConcentrations.
Bar-Ilan,O.etal.(2013)EnvironSciTechnol.47(9):4726-4733.
PubMedID:23510150ReactivityZebrafishApplications:DNAextract
PoorsleepinPCOS;ismelatonintheculprit?
Shreeve,N.etal.(2013)HumReprod.28(5):1348-1353.
PubMedID:23438443ReactivityHumanApplications:Urine
Grasshoppers"adaptationtoelevatedradioactivityintheChernobylexclusionzone.
Mortensen,L.H.(2013)RoskildeUniversityDissertation
PubMedID:N/AReactivityGrasshopperApplications:Intestines
ConsumptionofalowglycaemicindexdietinlatelifeextendsLifespanofBalb/cmicewithdifferentialeffectsonDNAdamage.
Nankervis,S.A.,Mitchell,J.M.,Charchar,F.J.,McGlynn,M.A.andLewandowski,P.A.(2013)LongevHealthspan.2(1):4.
PubMedID:24472560ReactivityMouseApplications:WBC"s
Light-inducedcelldetachmentforcellsheettechnology.
Hong,Y.etal.(2013)Biomaterials.34(1):11-18.
PubMedID:23069710ReactivityMouseApplications:MC3T3-E1cells
Puerarinpreventshighglucose-inducedapoptosisofSchwanncellsbyinhibitingoxidativestress.
Wu,Y.,Xue,B.,Li,X.andLiu,H.(2012)NeuralRegenRes.7(33):2583-2591.
PubMedID:25368634ReactivityMouseApplications:Schwanncells
TheprotectiveeffectofAlphalipoicacidonSchwanncellsexposedtoconstantorintermittenthighglucose.
Sun,L.etal.(2012)BiochemPharmacol.84(7):961-973.
PubMedID:22796564ReactivityMouseApplications:Schwanncells
GinsenosideRb1relievesglucosefluctuationinducedoxidativestressandapoptosisinSchwanncells.
Xue,B.etal.(2012)NeuralRegenRes.2012(30):2340-2346.
PubMedID:25538758ReactivityMouseApplications:Schwanncells
EicosapentaenoicAcidandOxypurinolintheTreatmentofMuscleWastinginaMouseModelofCancerCachexia.
Vaughan,V.C.,Sullivan-Gunn,M.,Hinch,E.,Martin,P.,Lewandowski,P.A.(2012)PLoSONE.7(9):e45900.
PubMedID:23029301ReactivityMouseApplications:Gastrocnemiusmuscle
Titaniumdioxidenanoparticlesproducephototoxicityinthedevelopingzebrafish.
Bar-Ilan,O.etal.(2012)Nanotoxicology.6(6):670-679.
PubMedID:21830861ReactivityZebrafishApplications:Embryos
EvaluationofvitaminB12effectsonDNAdamageinducedbypioglitazone.
Alzoubi,K.,Khabour,O.,Hussain,N.,Al-azzam,S.andMhaidat,N.(2012)MutatRes.748(1-2):48-51.
PubMedID:22790087ReactivityHumanApplications:Blood
ProtectiveEffectsofSalvianolicAcidBonSchwannCellsApoptosisInducedbyHighGlucose.
Sun,L.etal.(2012)NeuRochemicalRes.37(5):996-1010.
PubMedID:22252725ReactivityMouseApplications:Schwanncells
OxidativeStressinHPV-DrivenViralCarcinogenesis:RedoxProteomicsAnalysisofHPV-16DysplasticandNeoplasticTissues.
DeMarco,F.etal.(2012)PLoSONE.7(3):e34366.
PubMedID:22470562ReactivityHumanApplications:Ecto-cervicaltissue
InhibitoryeffectsofSalvianolicacidBonapoptosisofSchwanncellsanditsmechanisminducedbyintermittenthighglucose.
Sun,L.etal.(2012)LifeSci.90(3-4):99-108.
PubMedID:22036624ReactivityMouseApplications:Schwanncells
AqPCR-basedassaytoquantifyoxidizedguanineandotherFPG-sensitivebaselesionswithintelomericDNA.
O"Callaghan,N.,Baack,N.,Sharif,R.andFrench,M.(2011)Biotechniques.51(6):403-411.
PubMedID:22150331ReactivityHumanApplications:SpleenBlymphoblastoidcells
Theeffectofcocoasupplementationonhepaticsteatosis,reactiveoxygenspeciesandLFABPinaratmodelofNASH.
Janevski,M.,Antonas,K.N.,Sullivan-Gunn,M.J.,McGlynn,M.A.andLewandowski,P.A.(2011)CompHepatol.10(1):10.
PubMedID:22081873ReactivityRatApplications:Livertissue
ReductionofOxidativeDamageReflectsaBetterKidneyTransplantationOutcome.
LaManna,G. etal. (2011)AmJNephrol.34,496-504.
PubMedID:22041478ReactivityHumanApplications:PBMC
Integratedexposureassessmentofsewageworkerstogenotoxicants:anurinarybiomarkerapproachandoxidativestressevaluation.
Zabadi,H.A.etal.(2011)EnvironHealth.10(23).
PubMedID:21435260ReactivityHumanApplications:Urine
TransgenicMiceExpressingCyclooxygenase-2inHepatocytesRevealaMinorContributionofThisEnzymetoChemicalHepatocarcinogenesis.
Izquierdo,C.L.etal.(2011)AmJPathol.178(3):1361-1373.
PubMedID:21356386ReactivityMouseApplications:COX-2Tgcells
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
配制分三步： 1) 1 M Tris-HCl (pH 8.0) 50 ml的配制：称取Tris碱6.06 g，加超纯水40 ml溶解，滴加浓HCl约2.1 ml调pH至8.0，定容至50 ml。
电泳缓冲液的另一个作用是使溶液具有一定的导电性，以利于DNA分子的迁移，例如，一般电泳缓冲液中应含有0.01-0.04mol/L的Na+离子，Na+离子的浓度太低时电泳速度变慢；太高时就会造成过大的电流使胶发热甚至熔化。
电泳缓冲液还有一个组分是EDTA，加入浓度为1-2mmol/L，目的是螯合Mg2+ 等离子，防止电泳时激活 DNA酶，此外还可防止Mg2+离子与核酸生成沉淀。
