SDS–PAGE
and immunoblot examination were performed following the procedure
described previously.53 (link)−55 (link) Briefly, 20 or 40 μg of total protein was
loaded into each well for immunoblotting and Coomassie staining, respectively.
Immunoblot analysis was performed using primary mouse monoclonal anti-His
(Invitrogen, catalog no. MA1-135 dilution 1:3000), rabbit polyclonal
anti-CsoS1 (Agrisera, catalog no. AS142760, dilution 1:3000), and
horseradish peroxidase-conjugated goat anti-mouse IgG secondary antibody
(Agrisera, catalog no. AS111772, dilution 1:10 000) and anti-rabbit
IgG secondary antibody (Agrisera, catalog no. AS09602, dilution 1:10 000).
Signals were visualized by using a chemiluminescence kit (Bio-Rad).
Immunoblot images were collected by ImageQuant LAS 4000 software,
version 1.2.1.119. Immunoblot protein quantification was performed
using ImageJ software (version 1.52 h). For each experiment, at least
three biological repeats were examined.
and immunoblot examination were performed following the procedure
described previously.53 (link)−55 (link) Briefly, 20 or 40 μg of total protein was
loaded into each well for immunoblotting and Coomassie staining, respectively.
Immunoblot analysis was performed using primary mouse monoclonal anti-His
(Invitrogen, catalog no. MA1-135 dilution 1:3000), rabbit polyclonal
anti-CsoS1 (Agrisera, catalog no. AS142760, dilution 1:3000), and
horseradish peroxidase-conjugated goat anti-mouse IgG secondary antibody
(Agrisera, catalog no. AS111772, dilution 1:10 000) and anti-rabbit
IgG secondary antibody (Agrisera, catalog no. AS09602, dilution 1:10 000).
Signals were visualized by using a chemiluminescence kit (Bio-Rad).
Immunoblot images were collected by ImageQuant LAS 4000 software,
version 1.2.1.119. Immunoblot protein quantification was performed
using ImageJ software (version 1.52 h). For each experiment, at least
three biological repeats were examined.