Rabbit monoclonal anti bax antibody

Brain paraffin sections (4μm) were incubated overnight at 4°C with rabbit polyclonal anti-Bcl-2 antibody (1:200, Abcam, Cambridge, UK) and rabbit monoclonal anti-Bax antibody (1:200, Abcam, Cambridge, UK) respectively. After three times rinses (5 min each) with phosphate-buffered saline (PBS, pH=7.4), the sections reacted with goat anti-rabbit IgG conjugated to peroxidase secondary antibody (1:200, Aspen Biotechnology, Wuhan, China). The remaining procedures were according to the standard procedures [22] . Negative controls were established to investigate the specificity of the reactions. For semi-quantitative analysis, four visual fields (200 ×) from each section in periinfarct area were photographed under a light microscope (Olympus Corporation, Tokyo, Japan), and integrated optical density (IOD) was analyzed using Image Pro Plus 6.0 (Media Cybernetics Inc., Bethesda, MD, USA).
The immunofluorescent staining was used to assess the expression of caspase-3. Brain sections were then incubated overnight with rabbit polyclonal anti-caspase-3 antibody (1:200, Abcam, Cambridge, UK). After being rinsed with PBS, the tissues were incubated with Cy3-conjugated goat anti-rabbit IgG secondary antibody (1:50, Aspen Biotechnology, Wuhan, China). The unspecific labeling was found by replacing the primary antibody with PBS.