Description
Key facts
Detection method:Fluorescent,
Sample types:Suspension cells, Adherent cells,
Assay Platform:Flow cytometer
Product details:
Intracellular glutathione (GSH) Detection Assay Kit (ab112132) uses our proprietary non-fluorescent Green Dye, which becomes strongly fluorescent upon reacting with thiol (including GSH in cells). In normal cells, the Green Dye is accumulated primarily in cytosol, but it is partially translocated to mitochondria in apoptotic cells while Green Dye staining intensity is decreased. Cells stained with Green Dye can be visualized with a flow cytometer at Ex/Em = 490/520 nm (FL1 channel).
This kit has been designed to detect apoptosis is cells by measuring the decrease in reduced glutathione (GSH), which is an early hallmark in the progression of cell death in response to different apoptotic stimuli in many cells.
This kit can be used together with other reagents, such as 7-AAD for multi-parametric study of cell viability and apoptosis. The kit is optimized for screening apoptosis activators and inhibitors with a flow cytometer.
We do not recommend using this kit in a microplate. A more suitable option might be
ab65322
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Glutathione is a tripeptide that contains L-cysteine, L-glutamic acid and glycine. It is the smallest intracellular protein thiol molecule in the cells, which prevents cell damage caused by reactive oxygen species such as free radicals and peroxides. Glutathione exists in reduced (GSH) and oxidized (GSSG) states. Reduced glutathione (GSH) is a major tissue antioxidant that provides reducing equivalents for the glutathione peroxidase (GPx) catalyzed reduction of lipid hydroperoxides to their corresponding alcohols and hydrogen peroxide to water. In the GPx catalyzed reaction, the formation of a disulfide bond between two GSH molecules generates oxidized glutathione (GSSG). The enzyme glutathione reductase (GR) recycles GSSG to GSH with the simultaneous oxidation of β-nicotinamide adenine dinucleotide phosphate (β-NADPH2). In healthy cells, more than 90% of the total glutathione pool is in the reduced form (GSH). When cells are exposed to increased levels of oxidative stress, GSSG accumulates and the ratio of GSSG to GSH increases. An increased ratio of GSSG-to-GSH is an indication of oxidative stress. The monitoring of reduced and oxidized GSH in biological samples is essential for evaluating the redox and detoxification status of the cells and tissues against oxidative and free radicals mediated cell injury.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Glutathione often referred to as GSH is a small tripeptide with a molecular mass of approximately 307.33 g/mol. It is composed of amino acids glutamine cysteine and glycine. Glutathione is found in almost all cells with high concentrations especially in the liver. As a powerful antioxidant it functions by reducing reactive oxygen species (ROS) and maintaining the redox status of cells. Glutathione exists in two forms: reduced (GSH) and oxidized (GSSG). The balance between these forms known as the GSH/GSSG ratio is critical for cellular health and is commonly assessed using a glutathione assay kit.
Biological function summary
Glutathione plays an important role in detoxification processes protecting cells from damage caused by free radicals peroxides and heavy metals. It acts as a substrate for various enzymes like glutathione peroxidase and glutathione S-transferase involved in neutralizing oxidative damage. Glutathione doesn't typically form a complex but it does participate in disulfide bond exchange reactions which are critical in maintaining cellular protein functions and structures.
Pathways
Glutathione is integral to cellular antioxidant defense and phase II detoxification pathways. It closely interacts with proteins like glutathione peroxidase in the reduction of hydrogen peroxide to water. Also glutathione's role in the pentose phosphate pathway supports the regeneration of NADPH which is essential for maintaining the reduced form of glutathione. This interplay is important for protecting cells against oxidative stress and ensuring cellular energy is balanced.
Glutathione has significant implications in conditions such as Parkinson's disease and liver cirrhosis. In Parkinson's disease researchers observe lower levels of glutathione in affected brain regions suggesting an important role in neurodegeneration. Glutathione levels also impact liver health with deficiencies leading to ineffective detoxification and the potential for liver damage or cirrhosis. In these contexts glutathione interacts with proteins involved in redox balance and neurotransmitter regulation emphasizing its broad significance in maintaining human health.
