From these, we selected for estimation parameters that were not available from literature and that are most directly associated with the dynamics of the GSH and GSSG concentrations ([GSH] and [GSSG], respectively), for which we had available experimental data
The tissue homogenates were centrifuged for 12000 x g for 15 min and collected and diluted to 1 mg tissue protein per ml (1 mg/ml) in homogenization buffer (PBS, pH 7.4, containing a protease inhibitor cocktail) as the stock solution
Phentermine for Weight Loss Phentermine alone is one of the oldest and most widely prescribed weight loss medications in the United States
Immunofluorescence staining of SIOs SIOs were fixed in 4% paraformaldehyde (Biosharp, Hefei, China) at room temperature for 20 min, washed three times with PBS, blocked with blocking buffer at room temperature for 1 h, incubated overnight with a primary antibody at 4 C, washed three times with PBS, incubated with a secondary antibody at room temperature for 90 min, washed three times with PBS, stained with 4,6-diamidino-2-phenylindole (Sigma-Aldrich Corporation) for 10 min, washed three times with ultrapure water, mounted on glass slides, and observed with a laser confocal microscope (Leica Microsystems GmbH, Wetzlar, Germany)
(PubMed) MacLennan SC, Wade FM, Forrest KM, Ratanayake PD, Fagan E, Antony J
doi: 10.3389/fphar.2023.1196413, 89 MendozaE.ScharffC
