Westernbright ecl substrate kit

Protein expressions of Bax, Bcl-2, caspase-3, and p53 in the BF of broilers were estimated by western blot according to our previous study [51 (link)]. The following antibodies used for the present study were purchased from indicated sources: Anti-Bax, Bcl-2, caspase-3, and p53 (Abclonal Technology, Wuhan, China), and Anti-β-Actin, (Cell Signaling Technology, Boston, MA, USA). The HRP-labeled goat anti-rabbit IgG (Servicebio Technology, Wuhan, China) was used as the secondary antibody. Samples were analyzed in triplicate; a representative blot is shown in respective figures. Proteins bands were detected through chemiluminescence WesternBright^TM ECL substrate kit (Advansta, Menlo Park, CA, USA), then visualized and quantified by a FluroChem FC2 Imaging System (ProteinSimple, California, USA).

Protein expressions of Nrf2, HO-1, GPx1, NQO1 and GCLC in the liver and NF-κB, p-NF-κB, and IκBα in the spleen of broilers were determined by western blot according to our previous study [58 (link)]. The following antibodies used for the present study were purchased from indicated sources: Anti-NF-κB, Anti-p-NF-κB, Anti-Nrf2, Anti-HO-1, anti-GPx1, (Abcam, Cambridge, MA, USA), Anti-NQO1, GCLC (Abclonal Technology, Woburn, MA, USA), Anti-β-Actin, Anti-IκBα (Cell Signaling Technology, Boston, MA, USA). The HRP-labeled goat anti-rabbit IgG (Servicebio Technology, Wuhan, China) was used as the secondary antibody. Samples were analyzed in triplicate; a representative blot is shown in respective figures. The proteins bands were detected via chemiluminescence WesternBright^TM ECL substrate kit (Advansta, Menlo Park, CA, USA), then visualized and quantified by FluroChem FC2 Imaging System.