Rabbit anti bax

The lysate (20 μl) containing 20 μg of protein was resolved on SDS-PAGE electrophoresis. After subsequent transblotting (wet transfer), Polyvinylidene fluoride membrane (PVDF) was incubated in Tris-buffered saline (with 0.01% Tween 20) (TBST) for 15 minutes at room temperature. Thereafter, the membrane was blocked in 3% Bovine serum albumin (prepared in TBST) for 50 mins at room temperature. The protein of interest, and control (GAPDH) were detected using the following primary antibodies; Mouse anti- TH (Cell Signaling; Danvers, MA, USA), Rabbit anti- BDNF (PeproTECH; New Jersey, USA), Rabbit anti- MAO-B (Abcam; Cambridge, UK), Rabbit anti- DDC (Cell Signaling), Rabbit anti- DAT (abcam), Rabbit anti-GAPDH (Cell Signaling), Rabbit anti- COMT (Proteintech, Illinois, USA), Rabbit anti-BAX (Cell Signaling), Mouse anti- CD11b (Abcam), Rabbit anti- IL-1β (Cell Signaling) and Rabbit anti- p47phox (Enzo life Sci; New York, USA). All primary antibodies were diluted in the blocking solution at 1: 500–1,000. The primary antibodies were detected using HRP-conjugated secondary antibodies (goat anti-rabbit and goat anti-mouse; Invitrogen (California, USA); dilution of 1: 5,000–10,000). The membrane was scanned using Chemidoc scanner (Bio-Rad laboratories; California, USA) to reveal the protein bands, which were quantified using Bio-Rad’s image lab software.