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ARBOR ASSAYS/Glutathione (GSH) Fluorescent 384-Well Detection Kit/K006-F1D/F1D
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- MostSensitive
- MultiSpecies
- USE-MeasureFreeorOxidizedGlutathionein15Minutes.
- SAMPLE-Lysates,WB,RBCs,Serum,Plasma,UrineandTissue
- SAMPLES/KIT-~183(FreeandOxidized)inDuplicate
- STABILITY-Stable4˚CLiquidReagents
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TheDetectX®Glutathione(GSH)FluorescentDetectionkitutilizesaproprietarynon-fluorescentmolecule,ThioStar®,thatcovalentlybindstothefreethiolgrouponGSHtoyieldahighlyfluorescentproduct.AftermixingthesampleorstandardwithThioStar®andincubatingatroomtemperaturefor15minutes.Thiskituses2384-wellplatesthatallowthemeasurementofabout375samplesinduplicateforeitherGSHorGSSGcontent,ormeasureabout183samplesinduplicateforbothGSHandGSSG.Thesignalproducedisreadat510nminafluorescentplatereaderwithexcitationat390nm.SampleGSSGisdeterminedbyadditionofareactionmixtureconvertsalltheGSSGintofreeGSH,whichthenreactswithThioStar®.
Glutathione(GSH)isthehighestconcentrationnon-proteinthiolinmammaliancellsandispresentinconcentrationsof0.5to10mM.GSHplaysakeyroleinmanyBIOLOGicalprocesses,includingthesynthesisofproteinsandDNA,thetransportofaminoacids,andtheprotectionofcellsagainstoxidation.Harmfulhydrogenperoxidecellularlevelsareminimizedbytheenzymeglutathioneperoxidase(GP)usingGSHasareductant.TheoxidizedGSHdimer,GSSG,isformedfromGSHandperoxidebytheGPreaction.AnimportantroleofGSSGintheNFkBactivatingsignalcascadeissuggestedbythefactthatthepotentNFkBinducerTPAincreasesintracellularGSSGlevelsandGSSG/GSHratios.
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