Anti aldh2 mouse polyclonal antibody

Western blot was performed as described earlier^{29 (link)}. In brief, whole cardiac tissue was homogenized and then the tissue samples were separated on SDS-polyacrylamide gels by electrophoresis and were transferred to immobilon-P membranes (Millipore Sigma, ABN294). The levels of each protein were determined using specific antibodies: anti-ALDH2 mouse polyclonal antibody (Thermofisher Scientific Inc, MA5-17029); anti-4HNE-Cys/His/Lys rabbit polyclonal antibody (Millipore Sigma, 303207) and anti-β actin mouse monoclonal antibody (Abcam, sc-47778). All the antibodies are used at a concentration of 1:1000. The bound antibody was visualized with horseradish peroxidase (HRP)-coupled secondary antibody. The band intensities were quantified using Image J software.

Frozen cardiac tissue sections were used for immunostaining to determine the transfection efficiency by checking the eGFP florescence in mice received AAV9-ALDH2-eGFP and AAV9-GFP and as well as checking the staining for ALDH2 with anti-ALDH2 mouse polyclonal antibody (Thermo Fisher Scientific Inc, MA5-17029; 1:100 and 4 °C for overnight incubation): The secondary antibodies were conjugated with Alexa fluor 546 (Invitrogen, A11030, wavelength excitation peaks at 556 nm and emission peaks at 573 nm) at a concentration of 1:500 at room temperature for 1 hour. Immunofluorescence positive staining was analyzed using a fluorescent microscope (Olympus IX81) and an image analyzer (Olympus IX2-UCB). The representative micrographs were selected and presented.