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Abcam's Glutathione Detection Assay Kit (Fluorometric) provides a simple in vitro assay for detection of total glutathione changes in apoptosis and other samples. The assay utilizes monochlorobimane (MCB), a dye that appears to form an adduct exclusively with glutathione. The unbound MCB is almost nonfluorescent, whereas the dye fluoresces blue (Ex/Em = 380nm/461nm) when bound to glutathione of reduced or oxidized form. The reaction is catalyzed by glutathione S-transferase. Thus, the amount of total glutathione can be easily detected using a fluorometer or a 96-well fluorometric plate reader.
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Glutathione is the principal intracellular low-molecular-weight thiol that plays a critical role in the cellular defense against oxidative and nitrosative stress in mammalian cells. Diminished glutathione levels have been observed in the early stages of apoptosis.
|Cell Lysis Buffer||1 x 25ml|
|GSH Standard||Yellow||1 vial|
|GST Reagent||Green||1 x 200µl|
|Monochlorobimane Substrate||Red||1 x 200µl|
Our Abpromise guarantee covers the use of ab65322 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Functional Studies||Use at an assay dependent dilution.|
uninfected cells;WT: infected with M.tuberculosis wild type;
KO: infected with M.tuberculosis OppD knock-out;COM: infected with M.tuberculosis OppD knock-out complemented with OppDA gene.
106 cells were infected and lysed by treating them with 100µl of ice cold lysis buffer. Cell lysate was diluted and mixed as described in the kit protocol. After 30 min incubation at 37C, fluorescence was measured at Ex=380nm/ Em=460nm. Results represent the means of ± S.D. of three determinations.
Image obtained from Dasgupta A. et al; PLoS One; 2010 Aug 17; 5(8): e12225.
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