ProductName | NitricOxideDetectionKit |
Description | Colorimetricdetectionofnitrateandnitrite |
SpeciesReactivity | SpeciesIndependent |
Platform | Microplate |
SampleTypes | Buffer,Celllysates,Plasma,Saliva,Serum,TissueCultureMedia,Urine,Water |
DetectionMethod | ColorimetricAssay |
AssayType | IndirectQuantitativeAssay |
Utility | ColorimetricassayusedtoquantitativelymeasureNitrateandNitritepresentinavarietyofsamples. |
Sensitivity | 2.63µMintheNitriteand1.02µMintheTotalNitricOxide |
AssayRange | 3.125-200µM |
Precision | IntraAssayPrecision:ThreesampleswerefurtherdilutedinAssayBufferandruninreplicatesof20inanassay.ThemeanandprecisionofthecalculatedNitriteorTotalNOconcentrationswere:Sample1(NitriteConcentration)-45.1µM,4.4%CV,Sample1(TotalNOConcentration)-70.5µM,6.8%CVSample2(NitriteConcentration)-73.3µM,9.1%CV,Sample1(TotalNOConcentration)-107.4µM,4.4%CVSample3(NitriteConcentration)-132.7µM,1.3%CV,Sample1(TotalNOConcentration)-157.8µM,1.8%CVInterAssayPrecision:ThreesampleswerefurtherdilutedinAssayBufferandruninduplicatesintwentyassaysrunovermultipledaysbythreeoperators.ThemeanandprecisionofthecalculatedNitriteorTotalNOconcentrationswere:Sample1(NitriteConcentration)-44.1µM,3.1%CV,Sample1(TotalNOConcentration)-68.8µM,7.4%CVSample2(NitriteConcentration)-66.4µM,4.0%CV,Sample1(TotalNOConcentration)-112.1µM,5.7%CVSample3(NitriteConcentration)-126.7µM,6.3%CV,Sample1(TotalNOConcentration)-154.48µM,4.1%CV |
IncubationTime | 30minutes |
NumberofSamples | 88samplesinduplicate |
OtherResources | KitBooklet,MSDS |
StorageTemperature | 4ºC | ||||||||||||||||||||||||||||||
ShippingTemperature | BlueIce | ||||||||||||||||||||||||||||||
ProductType | DetectionKits | ||||||||||||||||||||||||||||||
AssayOverview | TheNitricOxideDetectionKitisdesignedtoquantitativelymeasureNitrateandNitritepresentinavarietyofsamples.NitricOxidecontentisderivedfromthesumofNitrate(-NO3)andNitrite(-NO2).BothNitrateandNitritestandardsareprovidedtogeneratestandardcurvesfortheassayandallsamplesshouldbereadofftheappropriatestandardcurve.ForNitritedetection,samplesaremixedwiththeColorReagentsAandBandincubatedatroomtemperaturefor5minutes.Thecoloredproductisreadat550–570nm.TheconcentrationofNitriteinthesampleiscalculated,aftermakingasuitablecorrectionforanydilutionofthesample,usingsoftwareavailablewithmostplatereaders.TotalNitricOxidecontentismeasuredafterthesampleisincubatedwithNitrateReductaseandNADH.ThereductaseincombinationwithNADHreducesNitratetoNitrite.Aftera20minuteincubationatroomtemperature,ColorReagentsAandBareaddedandincubatedatroomtemperaturefor5minutes.ThecoloredproductisreadandcalculatedaswiththeNitritedeterminationabove.TheconcentrationofNitrateinthesampleiscalculatedbysubtractingthemeasuredNitriteconcentrationfromtheTotalNitricOxideconcentrationforthesample. | ||||||||||||||||||||||||||||||
KitOverview |
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CiteThisProduct | NitricOxideDetectionKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-212) |
AlternativeNames | Nitrogenmonoxide;Nitrogen(II)oxideDetectionKit |
ResearchAreas | CellSignaling,Neuroscience,Post-translationalModifications |
ScientificBackground | Nitricoxide(NO)isadiffusIBLe,transient,reactivemoleculethathasphysiologicaleffectsinthepicomolar-to-micromolarrange.Actingthroughsolubleguanylatecyclaseactivation,NOisanimportantphysiologicalregulatorofthecardiovascular,nervous,andimmunologicalsystems(1).NOisbio-availablebytworoutes.ItcanbeendogenouslygeneratedbyconstitutiveorinducedenzymeslikeNitricOxideSynthaseoritcanbeorallyingestedasnitrates/nitritesforrapiduptakeintocirculationandsubsequentconversion(2).Thereactivenatureofnitricoxideallowsittoactasacytotoxicfactorwhenreleasedduringanimmuneresponsebycellssuchasmacrophages.ThereactivityalsoallowsNOtobeeasilyconvertedtoatoxicrADIcalthatcanproducenitrosativedamagetocells,organellesandmoleculessuchasDNA.Nitrosaylationhowevercanbearegulatedpost-translationalmodificationincellsignaling(3).Thebalanceanddynamicsoftheregulatory/damagefacetsofNOaremajorforcesinmitochondrialsignalinganddysfunction(4).NOislinkednotonlytocoronaryheartdisease,endothelialdysfunctions,erectiledysfunction,andneurologicaldisorders,butalsodiabetes,chronicperiodontitis,autism,cancer,andassortedage-relateddiseases(5-9).ThephysicalpropertiesofNitricOxidemakeitchallengingfordirectdetectionmethods.However,colorimetricmethodscanbeappliedtomeasureitsstablebreak-downproductsnitrate(-NO3)andnitrite(-NO2)(10). |
References | 1.Moncada,SandEAHiggs.(1991)Eur.J.Clin.Invest.,21:361-374. 2.Kapil,V.etal.(2010)Heart,96:1703-1709. 3.Seth,DandStamler,JS.(2007)Curr.Opin.Chem.Biol.,15:1-8. 4.Eursalimsky,JDandMoncada,S.(2007)ATVB27:2524-2531. 5.Knott,ABandBossy-Wetzel,E.(2010)Diab.Obes.Metab.12(Suppl2):126-133. 6.VanDyke,K.etal.(2010)Ann.N.Y.Acad.Sci.1203:138-145. 7.Reher,VSG.etal.(2007)J.OralSci.49(4):271-276. 8.Sogut,S.etal.(2003)Clin.Chim.Acta331:111-117. 9.Balam.E.etal.(2002)Jpn.J.Clin.Oncol.32(5):162-166. 10.Moshage,H.(1997)Clin.Chem.43(4):553-556. |
TypicalStandardCurveforNitricOxideDetectionKitStressXpress®–SKT-212.AssayType:CoupledEnzymeAssay.DetectionMethod:ColorimetricAssay.AssayRange:3.125–200uM.
LinearitywasdeterminedbytakingtwohumanurinesampleswithknownNitriteandTotalNitricOxideConcentrationsandmixingthemingivenratios.Themeasuredconcentrationswerecomparedtotheexpectedvaluesbasedontheratiosused.
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