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苏州工业园区生物纳米园A4#216
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商品描述
Overview:
| ProductName | GlutathioneDetectionKit |
| Description | FluorescentdetectionoftotalGSHcontent |
| SpeciesReactivity | SpeciesIndependent |
| Platform | Microplate |
| SampleTypes | Celllysates,EDTAPlasma,Erythrocytes,HeparinPlasma,Serum,Tissue,Urine,WholeBlood |
| DetectionMethod | FluorometricAssay |
| AssayType | DirectQuantitativeAssay |
| Utility | FluorometricdetectionassaytomeasurethetotalGSHcontentinsamples. |
| Sensitivity | 45nMintheFreeGSHand48nMintheTotalGSH |
| AssayRange | 0.195-25uM |
| Precision | IntraAssayPrecision:TwoeachofSSAtreatedhumanurineandwholebloodsampleswerefurtherdilutedin1%SSASampleDiluentandruninreplicatesof20inanassay.ThemeanandprecisionofthecalculatedGSHconcentrationswere:Sample1(Free)-1.27µM,4.0%CV,Sample1(Total)-2.30µM,4.7%CVSample2(Free)-2.00µM,3.1%CV,Sample2(Total)-3.80µM,4.7%CVSample3(Free)-8.33µM,4.6%CV,Sample3(Total)-9.77µM,2.7%CVSample4(Free)-3.89µM,3.0%CV,Sample4(Total)-4.45µM,2.3%CVInterAssayPrecision:TwoeachofSSAtreatedhumanurineandbloodsampleswerefurtherdilutedin1%SSASampleDiluentandruninduplicatesintwentyassaysrunovermultipledaysbytwooperators.ThemeanandprecisionofthecalculatedGSHconcentrationswere:Sample1(Free)-1.30µM,8.6%CV,Sample1(Total)-2.40µM,9.3%CVSample2(Free)-1.83µM,14.7%CV,Sample2(Total)-3.57µM,10.0%CVSample3(Free)-9.38µM,6.0%CV,Sample3(Total)-11.67µM,6.0%CVSample4(Free)-4.89µM,7.2%CV,Sample4(Total)-5.89µM,8.0%CV |
| NumberofSamples | 39samplesinduplicate |
| OtherResources | KitBooklet,MSDS |
Properties
| StorageTemperature | 4ºC | |||||||||||||||||||||||||||
| ShippingTemperature | BlueIce | |||||||||||||||||||||||||||
| ProductType | DetectionKits | |||||||||||||||||||||||||||
| AssayOverview | TheGlutathioneFluorescentDetectionkitisdesignedtoquantitativelymeasureglutathione(GSH),andoxidizedglutathione(GSSG)presentinavarietyofsamples.Thekitisuniqueinthatbothfreeandoxidizedglutathionearedetectedinthesamewellinthemicrotiterplate.Noseparationorwashingisrequired.TotalglutathioneisthesumofGSSGplusGSH.AGSHstandardisprovidedtogenerateastandardcurvefortheassayandallsamplesshouldbereadoffthestandardcurve.Thekitutilizesaproprietarynon-fluorescentmolecule,StressXpress®DetectionReagent,thatwillcovalentlybindtothefreethiolgrouponGSHtoyieldahighlyfluorescentproduct.AftermixingthesampleorstandardwiththeDetectionReagentandincubatingatroomtemperaturefor15minutes,thefluorescentproductisreadat510nminafluorescentplatereaderwithexcitationat390nm.TheconcentrationoftheGSHinthesampleiscalculated,aftermakingasuitablecorrectionforanydilutionofthesample,usingsoftwareavailablewithmostfluorescenceplatereaders.Freeglutathione,GSH,isreadfirstafter15minutes,followedbyadditionofareactionmixturethatconvertsalltheoxidizedglutathione,GSSG,intofreeGSH,whichthenreactswiththeexcessDetectionReagenttoyieldthesignalrelatedtoTotalGSHcontent.ThetotalconcentrationofGSHgeneratedinthesampleiscalculatedfromthegeneratedsignal.Wehaveprovideda96wellplateformeasurementbutthisassayisadaptableforhigherdensityplateformats.TheendusershouldensurethattheirHTSblackplateissuitableforusewiththesereagentspriortorunningsamples. | |||||||||||||||||||||||||||
| KitOverview |
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| CiteThisProduct | GlutathioneDetectionKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-202) |
BIOLOGicalDescription
| AlternativeNames | γ-L-Glutamyl-L-cysteinylglycineDetectionKit(2S)-2-Amino-5-[[(2R)-1-(carboxymethylamino)-1-oxo-3-sulfanylpropan-2-yl]amino]-5-oxopentanoicacidDetectionKit |
| ResearchAreas | Cancer,OxidativeStress |
| ScientificBackground | Glutathione(L-γ-glutamyl-L-cysteinylglycine;GSH)isthehighestconcentrationnon-proteinthiolinmammaliancellsandispresentinconcentrationsof0.5-10mM(1).GSHplaysakeyroleinmanybiologicalprocesses,includingthesynthesisofproteinsandDNA,thetransportofaminoacids,andtheprotectionofcellsagainstoxidation.Harmfulhydrogenperoxidecellularlevelsareminimizedbytheenzymeglutathioneperoxidase(GP)usingGSHasareductant(2).TheoxidizedGSHdimer,GSSG,isformedfromGSHandperoxidebytheGPreaction(seebelow).AnimportantroleofGSSGintheNFγBactivatingsignalcascadeissuggestedbythefactsthatthepotentNFγBinducer,tetradecanoylphorbolacetate,increasesintracellularGSSGlevelsandGSSG/GSHratios(3).GlutathioneS-transferases(GST)areanimportantgroupofenzymesthatcatalyzethenucleophilicadditionofGSHtoelectrophiles.Theyareencodedby5genefamilies;4encodecytosolicGSTandoneencodesthemicrosomalformofGST.Theyhavebeenimplicatedinanumberofdiseases.InasthmaarachidonicacidisconvertedtounstableleukotrieneA4(LTA4).LTA4iseitherhydratedtoformLTB4oritisconjugatedtoGSHbyaGST,leukotrieneC4synthase,toformleukotrieneC4.LTC4anditsderivativeLTD4areimportantmoleculesinbronchialasthma.LeukotrieneC4synthaseisthereforeanimportanttherapeutictarget.IthasalsobeenshownthatincreasedexpressionofGSTscanleadtodrugresistance.Threeglutathioneadductsofthedrugmelphalan,usedtotreatovariancancerandmultiplemyeloma,havebeenisolatedfromreactionsinvolvinghumanmicrosomalGSTs. |
| References | 1.MeisterA.(1988)TrendsBiochemSci.13(5):185-8. 2.MeisterA.(1994)JBiolChem.269(13):9397-400. 3.DrögeW.,etal.(1994)FASEBJ.8(14):1131-8. |
ProductImages
TypicalStandardCurveofGlutathioneDetectionKitStressXpress®–SKT-202.AssayType:Direct.DetectionMethod:FluorometricAssay.AssayRange:0.195–25uM
LinearitywasdeterminedbytakinghumanRBCsattwodifferentconcentrationsandmixedintheratiosgivenbelow.Themeasuredconcentrationswerecomparedtotheexpectedvaluesbasedontheratiosused.
Wepurchasedandcomparedapopularcolorimetrictotalglutathioneassaykit(kitT)thatusesEllmansreagenttodetectfreeglutathioneinthesample.Initialexperimentsusedrandomhumanurinesamplesthatwereprocessedasdescribedineachkitbooklet.WithkitT,thevaluesobtainedforurineaftertherecommendedtreatmentwith4volumesof5%metaphosphoricacidandsubsequent10folddilutionwithassaybufferputallthevalueswellbelowtheloweststandard.However,theurinesamplesrunintheStressXpress®kitgaveTotalGSHvaluesbetween0.63and4.04?M.
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