Glutathione Detection Kit (Blue Fluorescence) (BN00735)

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SKU:
BN00735
  • Glutathione Detection Kit (Blue Fluorescence) (BN00735)
  • Glutathione Detection Kit (Blue Fluorescence) (BN00735)
  • Glutathione Detection Kit (Blue Fluorescence) (BN00735)
€503

Description

ELISA Kit Technical ManualMSDS

Glutathione Detection Kit (Blue Fluorescence)

Glutathione (GSH) represents major low molecular-weight free thiol in living cells. As the key antioxidant in mammalian cells, intracellular GSH forms rapidly eliminated conjugates with electrophilic xenobiotics, free radicals as well as hydroperoxides. Insufficiency of cellular GSH results in oxidative stress damage and mitochondrial degeneration. Diminished levels of glutathione are symptomatic of cell stress thus; it is advantageous to monitor distribution of the intracellular GSH as an indicator of early apoptosis and cell death. Assay Genie's Glutathione Detection Kit (Blue Fluorescence) utilizes Monochlorobimane (MCB), a non-fluorescent and cell-permeable dye, that forms highly fluorescent adducts with GSH (GSH-MCB) detectable at EX/EM= 380/465 ±20 nm respectively. The reaction is catalyzed by glutathione-S-transferase (GST) and the intensity of the fluorescence reflects the amount of intracellular GSH. Hence, our kit provides a useful tool for fast and easy evaluation of the GSH effectors directly in the living cells.

Figure: Effect of Staurosporine on GSH levels reflected by reduction in fluorescence intensity in HeLa cells. HeLa cells were seeded overnight at 5 x 10 5 of viable cells/well. The next day the cell culture was treated with 0.5 µg/ml of Staurosporine and returned to the incubator for the next 24 h. Levels of GSH in the living cells corresponding to the intensity of the signal were determined as described in the Assay Protocol. Fluorescence was measured directly in white opaque plates with clear bottoms used for cell culturing. (I) Comparison of GSH levels in control and apoptotic cells: Panel A: control, untreated cells with bright uniform GSH staining; Panel B: apoptotic cells with diminished blue fluorescence post 24 h treatment with Staurosporine. Dead cells are visualized by red staining of nuclear DNA with Propidium Iodide. (II) GSH Fluorescence Curve of Jurkat cells prepared for this particular assay. Detection limit for corresponds to about 65,000 of Jurkat cells per well. Your results may not be identical to these. A new curve must be obtained for each experiment and the cell line. (III) Reduced levels of fluorescence in apoptotic Jurkat cells. Decrease in RFU values between cells treated overnight with Staurosporine vs untreated positive control. Background values correspond to the cell culture without MCB treatment.

Key Information Description

Product SKU

BN00735

Size

100 assays

Detection Method

Fluorescence (Ex/Em = 380/460 nm)

Application

  • Rapid and sensitive detection of intracellular levels of GSH and activity of GST in live cells
  • Screening for compounds affecting intracellular GSH levels

Features and Benefits

  • Highly sensitive fluorometric method to measure intra-cellular Glutathione levels in response to a variety of biochemical stimuli
  • Simple & High throughput-adaptable
  • Reproducible, Quantitative tool for screening, studying, and characterizing compounds that affect Glutathione metabolism

Kit Components

  • Wash Buffer
  • Monochlorobimane (MCB)

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans

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