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蚂蚁淘在线 / 品牌中心 / StressMarq / StressMarq/StressXpress® Glutathione Reductase Activity Kit/SKT-204-96/96-well
商品描述
Overview:
| ProductName | GlutathioneReductaseActivityKit |
| Description | FluorometricdetectionofglutathionereductaseactivitybytheamountofGSHgeneratedfromthereductionofGSSG |
| SpeciesReactivity | Human |
| Platform | Microplate |
| SampleTypes | Celllysates,EDTAPlasma,Erythrocytes,HeparinPlasma,Serum |
| DetectionMethod | FluorometricAssay |
| AssayType | DirectEnzymeActivityAssay |
| Utility | FluorometricassayusedtodetermineglutathionereductaseactivitybytheamountofGSHgeneratedfromthereductionofGSSG,insamples. |
| Sensitivity | 0.009mU/ml |
| AssayRange | 0.156-5mU/ml |
| Precision | IntraAssayPrecision:FivenativesamplesdilutedinAssayBufferandruninreplicatesof16inanassay.ThemeanandprecisionofthecalculatedGRactivitieswere:Sample1-3.35mU/mL,2.7%CVSample2-2.38mU/mL,5.6%CVSample3-1.75mU/mL,3.9%CVSample4-0.56mU/mL,3.7%CVSample5-0.27mU/mL,3.6%CVInterAssayPrecision:FivenativesamplesweredilutedinAssayBufferandruninduplicatesintwenty-twoassaysrunovermultipledaysbyfouroperators.ThemeanandprecisionofthecalculatedGRactivitieswere:Sample1-3.35mU/mL,5.0%CVSample2-2.36mU/mL,12.6%CVSample3-1.64mU/mL,6.8%CVSample4-0.62mU/mL,10.67%CVSample5-0.27mU/mL,10.5%CV |
| IncubationTime | 20Minutes |
| NumberofSamples | 41samplesinduplicate |
| OtherResources | KitBooklet,MSDS |
Properties
| StorageTemperature | 4ºC | |||||||||||||||||||||||||||
| ShippingTemperature | BlueIce | |||||||||||||||||||||||||||
| ProductType | ActivityKits | |||||||||||||||||||||||||||
| AssayOverview | TheGlutathioneReductaseActivitykitisdesignedtoquantitativelymeasureglutathionereductase(GR)activityinavarietyofsamples.AGRstandardisprovidedtogenerateastandardcurvefortheassayandallsamplesshouldbereadoffthestandardcurve.Thekitutilizesaproprietarynon-fluorescentmoleculethatwillcovalentlybindtothefreethiolgrouponGSHgeneratedinthereductionofoxidizedglutathione(GSSG)toyieldahighlyfluorescentproduct.AftermixingthesampleorstandardwithStressXpress®DetectionReagentandincubatingatroomtemperature,thefluorescentproductisreadat510nminafluorescentplatereaderwithexcitationat390nm.Backgroundthiolcontentisreadfirstafter5minutes,followedbyadditionofGSSGandNADPHwhichusesthestandardorsampleGRtoconverttheoxidizedglutathione,GSSG,intofreeGSH,whichthenreactswiththeStressXpress®DetectionReagenttoyieldthesignalrelatedtoGRactivityTheactivityofGRinthesampleiscalculatedfromthegeneratedsignal.Wehaveprovideda96wellplateformeasurementbutthisassayisadaptableforhigherdensityplateformats.TheendusershouldensurethattheirHTSblackplateissuitableforusewiththesereagentspriortorunningsamples. | |||||||||||||||||||||||||||
| KitOverview |
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| CiteThisProduct | GlutathioneReductaseActivityKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-204) |
BIOLOGicalDescription
| AlternativeNames | γ-L-Glutamyl-L-cysteinylglycineActivityKit(2S)-2-Amino-5-[[(2R)-1-(carboxymethylamino)-1-oxo-3-sulfanylpropan-2-yl]amino]-5-oxopentanoicacidActivityKit |
| ResearchAreas | Cancer,OxidativeStress |
| ScientificBackground | Glutathionereductase(GR)playsanindirectbutessentialroleinthepreventionofoxidativedamagewithinthecellbyhelpingtomaintainappropriatelevelsofintracellularglutathione(GSH).GSH,inconjuctionwiththeenzymeglutathioneperoxidase(GP),istheactingreductantresponsIBLeforminimizingharmfulhydrogenperoxidecellularlevels(1).TheregenerationofGSHiscatalyzedbyGR(2).GRisanubiquitous100-120kDadimericflavoproteinthatcatalyzesthereductionofoxidizedglutathione(GSSG)toreducedglutathione,usingß-nicotinamidedinucleotidephosphate(NADPH)asthehydrogendonor(3).MoleculessuchasNADPHactashydridedonorsinavarietyofenzymaticprocesses.NADPHhasbeensuggestedtoalsoactasanindirectlyoperatingantioxidant,givenitsroleinthere-reductionofGSSGtoGSHandthusmaintainingtheantioxidativepowerofglutathione. |
| References | 1.MeisterA.(1994)JBiolChem.269(13):9397-400. 2.MasseyV.,&WilliamsC.(1965)JBiolChem.240(11):4470-80. 3.AndersenH.,NielsenJ.,NielsenF.,&GrandjeanP.(1997)ClinChem.43(4):562-8. |
ProductImages
ContinuousKineticAssayTypicalStandardCurveforGlutathioneReductaseActivityKit(EnzymeActivityAssay)StressXpress®–SKT-204.AssayType:Direct.DetectionMethod:FluorometricAssay.AssayRange:0.156–5mU/ml.
End-PointAssayTypicalStandardCurveforGlutathioneReductaseActivityKit(EnzymeActivityAssay)StressXpress®–SKT-204.AssayType:Direct.DetectionMethod:FluorometricAssay.AssayRange:0.156–5mU/ml.
ChemicalequationoftheGlutathioneReductasereaction.TheregenerationofGSHiscatalyzedbyGlutathioneReductase.GlutathioneReductaseisanubiquitous100-120kDadimericflavoproteinthatcatalyzesthereductionofoxidizedglutathione(GSSG)toreducedglutathione,usingß-nicotinamidedinucleotidephosphate(NADPH)asthehydrogendonor.
LinearitywasdeterminedbytakingJurkatcelllysatesat40x106cells/mLdilutedto200,000and20,000cells/mLandmixingintheratiosgivenbelow.Themeasuredactivitieswerecomparedtotheexpectedvaluesbasedontheratiosused.
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