APT250 | Glutathione Detection Kit

APT250
100 assays  
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      Overview

      Replacement Information

      Key Spec Table

      Key ApplicationsDetection Methods
      ACTFluorescent
      Description
      Catalogue NumberAPT250
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionGlutathione Detection Kit
      OverviewReduced glutathione (GSH) plays a role in the rescue of cells from apoptosis, by buffering an endogenously induced oxidative stress. Maintaining intracellular glutathione levels can prevent apoptosis. Glutathione levels diminish with age. Intracellular levels of glutathione (reduced GSH) are directly influenced by the presence of extracellular glutathione (oxidized GSSG). Apoptotic cells extrude glutathione in the reduced form at an early stage of the apoptotic process; for example, Jurkat, U937 and Hep G2 cells are known to exhibit this change (Ghibelli et al. 1998). Evidence has accumulated that extrusion of reduced glutathione alters the intracellular redox state, and this change may trigger the loss of mitochondrial membrane potential (Hall 1999). However, some cells, like HeLa and 3T3, apparently do not undergo cytosolic GSH change during apoptosis. The glutathione pathway is also involved in drug resistance of certain cancer types such as leukemia (Sargent et al. 1999).

      Test Principle:

      After inducing apoptosis, incubate cell lysates with monochlorobimane (MCB), a dye that has a high affinity for glutathione (MCB binds other thiols in the cell with lower affinity). The unbound dye is almost nonfluorescent, whereas the thiol-bound dye fluoresces blue (excitation = 380 nm; emission = 461 nm). Fluorescence can easily be detected using a fluorometer or a 96-well fluorometric plate reader.

      Application:

      The CHEMICON Glutathione Detection Kit is a quick method for detection of intracellular levels of reduced glutathione (GSH). Reduction in levels of GSH may be an early indicator of apoptosis.
      Materials Required but Not Delivered· Microcentrifuge

      · Acetonitrile or DMF

      · Adjustable volume pipettor with disposable tips

      · Beakers, flasks, cylinders necessary for preparation of reagents

      · Fluorescence plate reader with 380nm/460nm filter set
      References
      Product Information
      Components
      • MCB Fluorochrome (Part No. 90127): 5 vials, 50 μg each.
      • 4X Cell Lysis Buffer (Part No. 90130): One 16 mL bottle.
      • 10X Wash Buffer (Part No. 90126): One 100 mL bottle.
      • GSH Positive Control (Part No. 90146): 1 vial, 100 mg.
      Detection methodFluorescent
      Applications
      ApplicationThe Glutathione Detection Kit is a quick method for detection of intracellular levels of reduced glutathione (GSH). Reduction in levels of GSH may be an early indicator of apoptosis.
      Key Applications
      • Activity Assay
      Biological Information
      Species Reactivity
      • Mammals
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsStore kit materials at 4°C up to their expiration date.
      Packaging Information
      Material Size100 assays
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      References

      Reference overviewPub Med ID
      Glucose depletion activates mmu-miR-466h-5p expression through oxidative stress and inhibition of histone deacetylation.
      Druz, A; Betenbaugh, M; Shiloach, J
      Nucleic acids research  40  7291-302  2012

      Show Abstract
      22638581 22638581

      User Guides

      Title
      Glutathione Detection Kit