Rabbit anti bax antibody

Sections were dewaxed and rehydrated by conventional methods. After quenching endogenous peroxidase with 3% hydrogen peroxide for 10 min and blocking with normal goat serum for 10 min at 37°C, sections were incubated with rabbit anti-Bax antibody (1:200) and mouse anti-Bcl-2 antibody (1:200) (both from Santa Cruz Biotechnology, Inc.) at 4°C overnight. After washing in PBS, the sections were incubated with FITC-conjugated anti-rabbit IgG (1:500) and Cy3 conjugated anti-mouse IgG (1:200) (both from Beijing Cowin Biotech Co., Ltd., Beijing, China) for 1 h at room temperature in the dark. Images were captured at a magnification of ×200 for analysis. The mean fluorescence intensity (MFI) was measured, and expression levels of Bax and Bcl-2 were calculated as change of the percentage in MFI compared to the vehicle control mice.

Western blot tests were performed as previously described [23 (link)]. Brains were removed, and tissues from the ipsilateral/left cerebral cortex were dissected rapidly and were frozen in liquid nitrogen at -80℃. After sample preparation, equal amounts of protein samples (50 μg) were loaded onto each lane of SDS-PAGE gel. After electrophoresis, the samples were transferred onto a nitrocellulose membrane, which was blocked with a blocking solution for 2 h. The membrane was incubated at 4 °C overnight with the following primary antibody: rabbit anti-TGR5 (1:1000, Abcam), rabbit anti-SIRT3 (1:1000, Abcam), rabbit anti-BCL-2 antibody (1:500; Santa Cruz), rabbit anti-BAX antibody (1:500; Santa Cruz), rabbit anti-cleaved caspase-3 antibody (1:100; Santa Cruz). β-actin was used as an internal loading control. The secondary antibodies were all from Santa Cruz Biotechnology. Blot bands were quantified by densitometry using ImageJ software (ImageJ 1.4; NIH, Bethesda, MD).