Rabbit polyclonal anti bax antibody

Western blot analysis was used to detect expression of pro-apoptotic (AIF, Bax) and anti-apoptotic (Bcl-2, Bcl-xL) proteins as well as p-Akt and cleaved Caspase-3. Tissue samples from the ischemic cerebral hemispheres of all experimental and control groups were harvested at 6 (n = 6 per group) or 24 h (n = 6 per group) after reperfusion, as described previously by us⁴⁵ . PVDF membranes after protein transfer were incubated with primary antibodies including rabbit polyclonal anti-Phospho-Akt antibody (1:1000, Cell Signaling Technology), rabbit polyclonal anti-Bcl-2 antibody (1:200, Santa Cruz), mouse monoclonal anti-Bcl-xL antibody (1:200, Santa Cruz), mouse monoclonal anti-AIF antibody (1:4000, Santa Cruz), rabbit polyclonal anti-Bax antibody (1:500, Santa Cruz), and rabbit polyclonal anti-cleaved Caspase-3 antibody (1:1000, Cell Signaling Technology) at 4 °C for 24 h. Membranes were then incubated with the relevant secondary antibody (goat anti-mouse IgG, ZhongShanJinQiao; goat anti-rabbit IgG, ZhongShanJinQiao) for 1 h at room temperature. Equal protein loading was adjusted using β-actin (mouse polyclonal anti-β-actin antibody, Santa Cruz). An ECL system was used to detect immunoreactive bands by luminescence. Quantification of relative target protein expression was obtained using an image analysis program (Image J 1.48, National Institutes of Health, USA).