Film multi gauge image analyzer software

Mouse brains and cells were resuspended in lysis buffer (10 mM Tris-HCl, pH 7.5, containing a protease inhibitors cocktail and phosphatase inhibitors (1 mM sodium orthovanadate, 5 µM sodium fluoride) and then sonicated before western blot analysis. Expressions of proteins were analyzed with 50 µg of cell lines or mouse brain homogenates loaded on 10–12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and semi-dry transferred for 10 min by means of the ready to use transfer kit nitrocellulose (Bio-Rad, 1704271) and the Trans-Blot® Turbo™ Transfer System (pre-programmed Bio-Rad protocol for 2 mini gels of 1.5 mm). Transferred proteins were then immunoblotted using the antibodies listed in Table 1. The full gels of PINK1 containing migration controls were provided for each cell and tissue type (first time appearance) to illustrate antibody specificity. Immunological complexes were revealed with adequate anti-rabbit or anti-mouse IgG-coupled peroxidase antibodies (Jackson ImmunoResearch, 111–036-045) by the electrochemiluminescence detection method (Roche Diagnostics S.A.S). Chemiluminescence was recorded using a luminescence image analyzer LAS-4000 (Raytest, Fuji) and quantifications of non-saturated images were performed with the FUJI Film Multi Gauge image analyzer software.