Rabbit polyclonal anti erk1 2

For protein analysis, samples were run on 7.5 or 10 % native or sodium dodecyl sulfate–polyacrylamide gels (SDS-PAGE). For visualization of separated proteins, gel was stained with 0.1 % Coomassie Blue R250 in 10 % acetic acid, 50 % methanol and 40 % H₂O and destained with 10 % acetic acid, 50 % methanol and 40 % H₂O. From other gels proteins were transferred onto polyvinylidene fluoride membrane by semidry western blotting. For specific detection, the following primary antibodies were used: rabbit polyclonal anti A1AT (DAKO), mouse monoclonal anti A1AT (clone B9, Santa Cruz Biotechnology), mouse monoclonal anti polymeric A1AT (Clone 2C1, Hycult biotech), mouse monoclonal anti FABP4 (clone 1105CT1-1-1, Antibodies-Online GmbH), rabbit polyclonal anti ERK1/2 and mouse monoclonal anti p-ERK1/2 (both from Sigma-Aldrich), monoclonal anti β-actin (AC-15, Sigma-Aldrich). The immune-complexes were visualized with appropriate secondary horseradish peroxidase-conjugated antibodies (DAKO A/S) and ECL Western blotting substrate (Thermo Fisher Scientific). The density of the specific bands was quantified using ImageJ software (http://imagej.nih.gov/ij)