relevant concentrations  (MedChemExpress)

Journal: Virology Journal

Article Title: HIV-1 tat expression and sulphamethoxazole hydroxylamine mediated oxidative stress alter the disulfide proteome in Jurkat T cells

doi: 10.1186/s12985-018-0991-x

Figure Lengend Snippet: The effect of Tat expression and SMX-HA-mediated oxidative stress on Prx1 oxidation in Jurkat T cells. Cells from the various cell lines were induced with 0, 400 or 1000 ng/ml Dox for 48 h, treated with 0. 5% DMSO or 100μM SMX-HA for 2 h followed by another 2 h incubation in media. The cells were then collected, cell lysates prepared and 25μg protein were dissolved in non-reducing loading buffer. The protein samples were run on 15% SDS-PAGE and transferred to membranes that were then probed with antibodies to Prx1. a A representative immunoblot showing oxidized dimers (labelled D on blots) and hyperoxidized monomers (labelled M on blots). b Quantification of the Prx1 monomer:dimer ratio in the Tat-expressing cell lines, presented as means ± S.E

Article Snippet: The cells were treated with 200 μM SMX-HA (a pharmacologically relevant concentration [ , ]) or the vehicle (DMSO) (Millipore Sigma, Oakville, ON, Canada) at a final concentration of 0.5% then incubated at 37 °C for 2 h. The cells were subsequently collected and lysed in RIPA buffer containing 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1% Triton x-100, 1% sodium deoxycholate, 0.1% SDS, 1 mM EDTA (pH 8.0), complete mini protease inhibitor cocktail (Roche) and 40 mM iodoacetamide ([IA]; Millipore Sigma, Oakville, ON, Canada).

Techniques: Expressing, Incubation, SDS Page, Western Blot