Mouse monoclonal α sarcomeric actin antibody

We detected apoptosis using the Dead End Fluorometric TUNEL System (Promega), according to the manufacturer's instructions. Four frozen hearts per group were sliced into 5 μm thick sections using a freezing microtome. Frozen sections were counterstained with mouse monoclonal α-sarcomeric actin antibody (1 : 75, Abcam), and Alexa Fluor 594 rabbit anti-mouse antibody (1 : 1000, Invitrogen) was applied as secondary antibody; cell nuclei were stained with 4′,6-Diamidino-2-Phenylindole, Dilactate (DAPI; Sigma). The stained sections were mounted and analyzed with a Fluoview 1000 confocal microscope (Olympus, Japan). We counted 6 random 400x fields and the number of TUNEL-positive cardiomyocyte nuclei was manually determined. The total number of nuclei (exhibited as DAPI-positive signals) was automatically calculated using Image-Pro Plus (version 5.0).

Cellular apoptosis was evaluated using DeadEnd Fluorimetric TUNEL System (Roche) according to the manufacturer’s instructions. Paraffin-embedded tissue sections were counterstained with mouse monoclonal α-sarcomeric actin antibody (1:75, Abcam) and Alexa Fluor 633 goat anti-mouse antibody (1:250, Molecular Probes) as secondary antibody. DAPI as nuclei marker. Sections were mounted and analyzed using a Fluoview 1000 confocal microscope (Olympus, Japan). The number of TUNEL-positive cardiomyocytes nuclei was manually determined. The total number of nuclei (exhibited as DAPI-positive signals) was automatically calculated using Image Pro Plus software (Media Cybernetic).