Overview:
ProductName | HydrogenPeroxideDetectionKit |
Description | QuantitativecolorimetricmeasurementofH2O2 |
SpeciesReactivity | SpeciesIndependent |
Platform | Microplate |
SampleTypes | Buffer,TissueCultureMedia,Urine |
DetectionMethod | ColorimetricAssay |
AssayType | DirectQuantitativeAssay |
Utility | ColorimetricassayusedtoquantitativelymeasureH2O2inavarietyofsamples. |
Sensitivity | 1.83µM |
AssayRange | 3.125-100µM |
Precision | IntraAssayPrecision:Threebuffersampleswereruninreplicatesof20inanassay.Themeanandprecisionofthecalculatedconcentrationswere:Sample1-82.2µM,2.1%CVSample2-53.1µM,2.4%CVSample3-19.4µM,5.9%CVInterAssayPrecision:Threebuffersampleswereruninduplicateintwelveassaysrunovermultipledaysbythreeoperators.Themeanandprecisionofthecalculatedconcentrationswere:Sample1-79.9µM,3.7%CVSample2-49.5µM,4.5%CVSample3-18.4µM,4.3%CV |
IncubationTime | 15Minutes |
NumberofSamples | 89samplesinduplicate |
OtherResources | KitBooklet,MSDS |
Properties
StorageTemperature | 4ºC | ||||||||||||||||||
ShippingTemperature | BlueIce | ||||||||||||||||||
ProductType | DetectionKits | ||||||||||||||||||
AssayOverview | TheHydrogenPeroxideDetectionKitisdesignedtoquantitativelymeasureH2O2inavarietyofsamples.Ahydrogenperoxidestandardisprovidedtogenerateastandardcurvefortheassayandallsamplesshouldbereadoffthestandardcurve.SamplesaremixedwiththeColorimetricSubstrateandthereactioninitiatedbyadditionofhorserADIshperoxidase.Thereactionisincubatedatroomtemperaturefor15minutes.TheHRPreactswiththesubstrateinthepresenceofhydrogenperoxidetoconvertthecolorlesssubstrateintoacoloredproduct.Thepinkproductisreadat560nm.IncreasinglevelsofH2O2causealinearincreaseincolor. | ||||||||||||||||||
KitOverview |
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CiteThisProduct | HydrogenPeroxideDetectionKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-216) |
BIOLOGicalDescription
AlternativeNames | DioxidaneDetectionKit,OxidanylDetectionKit |
ResearchAreas | Cancer,CellSignaling,Oxidation,OxidativeStress,Post-translationalModifications |
ScientificBackground | InbiologicalsystemsincompletereductionofO2duringrespirationproducessuperoxideanion(O2-·),whichisspontaneouslyorenzymaticallydismutatedbysuperoxidedismutasetoH2O2.ManycellsproducelowlevelsofO2-·andH2O2inresponsetoavarietyofextracellularstimuli,suchascytokines(TGF-ß1,TNF-a,andvariousinterleukins),peptidegrowthfactors(PDGF;EGF,VEGF,bFGF,andinsulin),theagoNISTsofheterotrimericGprotein–coupledreceptors(GPCR)suchasangiotensinII,thrombin,lysophosphatidicacid,sphingosine1-phosphate,histamine,andbradykinin,andbyshearstress(1).TheadditionofexogenousH2O2ortheintracellularproductioninresponsetoreceptorstimulationaffectsthefunctionofvariousproteins,includingproteinkinases,proteinphosphatases,transcriptionfactors,phospholipases,ionchannels,andGproteins.In1894,Fenton(2)describedtheoxidationoftartaricacidbyFe2+andH2O2.H2O2andO2mayparticipateintheproductionofsingletoxygenandperoxynitriteandthegenerationofthesespeciesmaybeconcurrentwithreactionsinvolvingiron,andundersomecircumstancestheymightbeimportantcontributorstoH2O2toxicity(3,4).AsubstantialportionofH2O2lethalityinvolvesDNAdamagebyoxidantsgeneratedfromiron-mediatedFentonreactions(5,6).DamagebyFentonoxidantsoccursattheDNAbasesoratthesugarresidues.Sugardamageisinitiatedbyhydrogenabstractionfromoneofthedeoxyribosecarbons,andthepredominantconsequenceiseventualstrandbreakageandbaserelease(7,8). |
References | 1.RheeSG,BaeYS,LeeSR,KwonJ.(2000)Science’sstke.Availableat:http://stke.sciencemag.org/cgi/content/abstract/sigtrans;2000/53/pe1 2.Fenton,HJH.J.Chem.Soc.(Lond.)1894,65:899–910. 3.Sies,H.Mutat.Res.,1993,299:183–191. 4.Squadrito,GL.,andPryor,WA.(1995)Chem.Biol.Interact.96:203–206. 5.Imlay,JA.,andLinn,S.(1988)Science240:1302–1309. 6.Mello-Filho,AC.,Meneghini,R.(1991)Mutat.Res.,251:109–113. 7.vonSonntag,C.(1987)pp.238–249,TaylorandFrancis,NewYork. 8.Henle,ES.,Roots,R.,Holley,WR.,andChatterjee,A.(1995)Radiat.Res.143:144–150. |
ProductImages
TypicalStandardCurveforHydrogenPeroxideDetectionKitStressXpress®–SKT-216.AssayType:DirectEnzyme.DetectionMethod:ColorimetricAssay.AssayRange:3.125–100?M.
LinearitywasdeterminedbytakingtwodilutedhumanurinesampleswithknownH2O2concentrationsandmixingthemingivenratios.Themeasuredconcentrationswerecomparedtotheexpectedvaluesbasedontheratiosused.
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