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商品详细StressMarq/StressXpress®谷胱甘肽检测试剂盒/SKT-202-96/96井
StressMarq/StressXpress®谷胱甘肽检测试剂盒/SKT-202-96/96井
StressMarq/StressXpress®谷胱甘肽检测试剂盒/SKT-202-96/96井
商品编号: SKT-202-96
市场价: ¥7180.00
美元价: 4308.00
产地: 美国(厂家直采)
公司:
产品分类: 酸碱缓冲液
公司分类: acid_base_buffer_solution
联系Q Q: 3392242852
电话号码: 4000-520-616
电子邮箱: info@ebiomall.com
商品介绍

Overview:

ProductNameGlutathioneDetectionKit
Description

FluorescentdetectionoftotalGSHcontent

SpeciesReactivitySpeciesIndependent
PlatformMicroplate
SampleTypesCelllysates,EDTAPlasma,Erythrocytes,HeparinPlasma,Serum,Tissue,Urine,WholeBlood
DetectionMethodFluorometricAssay
AssayTypeDirectQuantitativeAssay
UtilityFluorometricdetectionassaytomeasurethetotalGSHcontentinsamples.
Sensitivity45nMintheFreeGSHand48nMintheTotalGSH
AssayRange0.195-25uM
PrecisionIntraAssayPrecision: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
NumberofSamples39samplesinduplicate
OtherResourcesKitBooklet,MSDS

Properties

StorageTemperature4ºC
ShippingTemperatureBlueIce
ProductTypeDetectionKits
AssayOverviewTheGlutathioneFluorescentDetectionkitisdesignedtoquantitativelymeasureglutathione(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
ComponentNo.ItemQuantity/Size
SKC-202ABlack96WellPlate1Each
SKC-202BGlutathioneStandard100µl
SKC-202CStressXpress®DetectionReagent2PlaticVials
SKC-202DDryDMSO4ml
SKC-202EAssayBuffer60ml
SKC-202FNADPHConcentrate300µl
SKC-202GGlutathioneReductaseConcentrate300µl
SKC-202HOxidizedGlutathioneControl300µl
CiteThisProductGlutathioneDetectionKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-202)

BIOLOGicalDescription

AlternativeNamesγ-L-Glutamyl-L-cysteinylglycineDetectionKit(2S)-2-Amino-5-[[(2R)-1-(carboxymethylamino)-1-oxo-3-sulfanylpropan-2-yl]amino]-5-oxopentanoicacidDetectionKit
ResearchAreasCancer,OxidativeStress
ScientificBackgroundGlutathione(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.
References1.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

<p>Typical Standard Curve of Glutathione Detection Kit StressXpress® – SKT-202. Assay Type: Direct. Detection Method: Fluorometric Assay. Assay Range: 0.195 – 25 uM</p>

TypicalStandardCurveofGlutathioneDetectionKitStressXpress®–SKT-202.AssayType:Direct.DetectionMethod:FluorometricAssay.AssayRange:0.195–25uM

<p>Linearity was determined by taking human RBCs at two different concentrations and mixed in the ratios given below. The measured concentrations were compared to the expected values based on the ratios used.</p>

LinearitywasdeterminedbytakinghumanRBCsattwodifferentconcentrationsandmixedintheratiosgivenbelow.Themeasuredconcentrationswerecomparedtotheexpectedvaluesbasedontheratiosused.

<p>We purchased and compared a popular colorimetric total glutathione assay kit (kit “T”) that uses Ellman’s reagent to detect free glutathione in the sample. Initial experiments used random human urine samples that were processed as described in each kit booklet. With kit “T”, the values obtained for urine after the recommended treatment with 4 volumes of 5% metaphosphoric acid and subsequent 10 fold dilution with assay buffer put all the values well below the lowest standard. However, the urine samples run in the StressXpress® kit gave Total GSH values between 0.63 and 4.04 ?M.</p>

Wepurchasedandcomparedapopularcolorimetrictotalglutathioneassaykit(kit“T”)thatusesEllman’sreagenttodetectfreeglutathioneinthesample.Initialexperimentsusedrandomhumanurinesamplesthatwereprocessedasdescribedineachkitbooklet.Withkit“T”,thevaluesobtainedforurineaftertherecommendedtreatmentwith4volumesof5%metaphosphoricacidandsubsequent10folddilutionwithassaybufferputallthevalueswellbelowtheloweststandard.However,theurinesamplesrunintheStressXpress®kitgaveTotalGSHvaluesbetween0.63and4.04?M.

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品牌介绍
StressMarq Biosciences公司的核心技术领域为细胞应激与离子通道以及载体研究,同时在其他领域也取得了一定成就,包括翻译后修饰,提供甲基化与乙酰基化抗体。其中,细胞应激领域主要包括热休克蛋白(HSP)领域。我们公司不仅在热休克蛋白领域领先全球,而且在氧化应激领域也卓有成就。StressMarq的优势在于提供四种独立的产品系列,分别涉及抗体、蛋白、酶联免疫吸附试验(ELISA)试剂盒及小分子领域。