Glutathione-Mediated Oxidative Stress Assay
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Corresponding Organization :
Other organizations : Case Western Reserve University
Protocol cited in 2 other protocols
Variable analysis
- Reduced L-glutathione
- Sodium borohydride
- 2-methyl-2-butene solution (2.0 M in THF)
- Sodium chlorite
- Tert-butanol
- Glutathione reductase from baker's yeast (250 units/ml. Sigma catalogue #G3664)
- 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB)
- NADPH
- Not explicitly mentioned
- HOHA-lactone (
1 ) was prepared as described previously. - N,O-Bis(trimethylsilyl)trifluoroacetamide (BSTFA) was obtained from Acros Organics (Thermo Fisher, New Jersey).
- Dulbecco's modified Eagle's medium, high glucose (DMEM/F12), fetal bovine serum, penicillin–streptomycin and L-glutamine were obtained from Gibco (Life Technologies, Grand Island, NY).
- ARPE-19 cells were obtained from American Type Culture Collection (ATCC, Rockville, MD).
- The hRPE cells were obtained from Lonza Walkersville, Inc., Walkersville, MD, catalog # 00194987, lot # 0000424472. According to the manufacturer, they exhibited pan-cytokeratin staining (95%), anti-fibroblast staining (0%) and ZO-1 staining (strongly positive), and they have viability >70%, average doubling time 22 hrs.
- Retinal pigmented epithelial cell basal medium, an optimized mixture of growth factors and supplements for primary hRPE cells (SingleQuots™ Kit), was obtained from Lonza (Allendale, NJ).
- Not explicitly mentioned
- Not explicitly mentioned
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