Glutathione (GSSG) level in Chang's liver cell

Value 1.3 nmol/mg protein
Organism Mammalian tissue culture cell
Reference Crespo et al., Differential effects of dietary flavonoids on reactive oxygen and nitrogen species generation and changes in antioxidant enzyme expression induced by proinflammatory cytokines in Chang Liver cells. Food Chem Toxicol. 2008 May46(5):1555-69. doi: 10.1016/j.fct.2007.12.014. p.1557 figure 2APubMed ID18234413
Method P.1557 right column bottom paragraph to p.1558 left column: "Cells were homogenised in 0.1 M sodium phosphate, 5 mM EDTA buffer (pH 8.0) with 25% phosphoric acid at a proportion of 1:20. The mixture was centrifuged at 100,000g for 30 min at 4 °C, the supernatant was collected and 500 µL were diluted with 4.5 mL of buffer. Two spectrophotometry cuvettes per sample were prepared with 1.8 mL phosphate-EDTA buffer, 100 µL supernatant and 100 µL O-phthalaldehyde (7.5 µM). After incubating for 15 min at 4 °C, a spectrofluorometric reading was obtained at an excitation wavelength of 350 nm and an emission wavelength of 420 nm. To find the percentage of glutathione corresponding to oxidised and reduced forms, 500 µL of the sample supernatant was incubated with 20 µL 4-vinylpyridine for 30 min to this mixture 4.5 mL of 0.1 M NaOH was added. A 100 µL portion of this mixture was then processed as described above to determine GSSG. GSH was obtained by subtracting GSSG from total glutathione." Value extracted visually from figure 2A
Comments For GSSG level in Chang's liver cell incubated with cytokine mixture see BNID 105115
Entered by Uri M
ID 102531