CA-G055 | |
Name | EliteTM Glutathione(GSH)AssayKit(Fluorescence) |
Description | TheEliteTMGlutathioneAssayKitprovidesanultrasensitivefluorimetricassaytoquantifyGSHinsample.Theproprietarynon-fluorescentglutathionesensorusedinthekitbecomesstronglygreenfluorescentuponreactingwithaGSHcompound,whichhasthespectralpropertiesalmostidenticaltothoseoffluoresceinandcanbeeasilyreadbyafluorescencemicroplatereaderatEx/Em=490/520nm.Thekitcandetectaslittleas1picomoleofGSHina100μLassayvolume(10nM).Inaddition,bothabsorptionandemissionspectraoftheglutathioneadductarepH-independent,makingthisassaykithighlyrobust. |
Application | Theassaycanbeperformedinaconvenient96-wellor384-wellmicrotiter-plateformatandeasilyadaptedtoautomationwithoutaseparationstep. |
Size | 200assays |
Detection | Fluorescencemicroplatereader |
Clickherefor DATASHEET | Clickherefor MSDS |
FrequentlyAskedQuestions:
1.Q:Whatwouldyousuggesttopreparethecellsamplesforthisassay?
Answer:Allthedetergentsinvariouslysisbufferswillcausebackgroundnoisefortheassay.ThetrADItionalwaytodoitisthe"freezeandthaw"method(i.e.freezefor20-30minutesat-80Cthenthawatroomtemperature,2-3times),butitistootedious.Wefoundthat0.2%SDSgivesthebestresultsamongallthedetergentstested.
2.Q:DoesthisKitworkforplantsamples?Couldyourecommendaspecialprotocoltomeasuregluthationeinplants?Answer:Aslongasthesamplchop2gofplantparts(leaf,root,grain,etc.)andgrounditinahomogenizerfor2minwith5-10mlof0.1NHCl;centrifugetopbtaintheGSHcontainingsupernatant,thenadjustpHto6.5fortheGSHassay.
3.Q:Mysampleisfrozenbraintissue,howshouldIprepareitforthesaasy?
Answer:ItisbetterusePBS,andsimplyhomogenizeit,thenusethesupernatant.
4.Q:Iworkwithhumanprimarycells.HowmanycellsshouldIuseintheassay?
Answer:Suggesttostartwith10,000cells/wellfirst.
5.Q:Forthefluorescentdetection,anemissionfilterwith520nmisrecommended.Butwehaveonly535nmfilter.Woulditworkforthismeasurement?
Answer:Yes.
6.Q:Wedon"thavethefiltersfor490/520(nm).Whatwehavearefor395/535(nm).Woulditwork?
Answer:No.theemissionisOK,buttheexcitationisnot.Thelowestexcitationwavelengthisat460nm.
7.Q: Couldyoutakealookatmyprotocolforpreparingthecellculturelysatefortheassayandgiveyoursuggestion?
Answer:
-removesupernatant,thenwashwithHEPES-BSS;[Fine]
-trypsinizefor2-6min,stoptrypsination;[WesuggesttoavoidtrypsinandtryusingarubberscrapperorEDTAtoharvestcells,thencentrifugethecellsandusethesupernatant]
-centrifugepelletdownat180g,5min,4°C,thenwashwith5mlcoldPBS;countcells;[Fine]
-centrifugepelletdownat180g,5min,4°C;[Mayneedtousemorecellssinceyouindicatedthesignalintheundilutedsampleisverylow.]
-washwith5mlCOLDPBS;resUSPendinice-cold5%MPA;[Fine.5%MPAcouldbeaddedtothesupernatantfordeproteination.]
-mixthoroughly,homogenizebysonication(2x5cycles),thencentrifugeat12.00rpmfor5minat4°C;[Fine]
-collectsupernatantforGSHassay;storeoniceORat-80°C.[NeedtoneutralizetheMPAtopH4~6,thenanalyzetheundilutedsamplewiththekit]
eENZYME为科学界生产和销售高质量的生物试剂盒和生物分析试剂盒。与NIH密切合作以保证产品在生命科学研究的多个领域成功地使用。努力提供新颖价廉且有效的工具，使实验更容易成功。产品在科研、生物技术、制药和政府的研究实验室中得到广泛的应用，包括药物发现、癌症研究、传染病研究、微生物学和分子诊断。
eENZYME is a manufacturer and distributor of high quality biological reagents and bio-assay kits to the scientific community. We work closely with the NIH and CDC communities to guarantee the successful performance of our products for life science research in broad areas. We strive to provide innovative and cost effective tools to make your success easier.
Our products are used in academic, biotechnology, pharmaceutical and government research laboratories in many diverse applications that include drug discovery, cancer research, infectious disease research, microbiology and molecular diagnostics.
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