Rabbit anti nf κb monoclonal antibody

Lung tissues were snap frozen in liquid nitrogen, pulverized and resuspended in ice-cold lysis buffer (Solarbio, Beijing, China). Protein concentrations were determined with the Bradford method. Lysates were allowed to solubilize on ice for 30 min, and particulate mass was removed by centrifugation at 15,000 × g for 15 min at 4°C. Supernatants were analyzed by SDS-PAGE. Primary antibodies used included rabbit anti-Tollip monoclonal antibody (1:400), rabbit anti-NF-κB monoclonal antibody (1:400), rabbit anti-IRAK1 monoclonal antibody (1:400), rabbit anti-TLR4 monoclonal antibody (1:400), rabbit anti-TRAF6 monoclonal antibody (1:400), rabbit anti-VEGF-α monoclonal antibody (1:400), rabbit anti-Nrf2 monoclonal antibody (1:400), mouse anti-GAPDH monoclonal antibody (1:400) were purchased from Santa Cruz Biotechnology, Inc.. Secondary antibodies were horseradish peroxidase-labeled antibodies (Thermo Scientific Pierce, Rockford, IL, USA). Blots were processed for enhanced chemifluorescence using a Pierce ECL Western blotting substrate (Thermo Scientific Pierce).

Lung tissues and isolated PBMCs were snap-frozen in liquid nitrogen, pulverized and resuspended in ice-cold lysis buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China). Protein concentrations were determined using the Bradford method (22 (link)). Lysates were solubilized on ice for 30 min and the particulate mass was removed using centrifugation (15,000 xg) for 15 min at 4°C. Supernatants were analyzed using 10% SDS-PAGE (Beijing Saichi Biological Technology Co., Ltd, Beijing, China). The primary antibodies used included rabbit anti-TIPE2 monoclonal antibody (1:400), rabbit anti-HO-1 monoclonal antibody (1:400), rabbit anti-NF-κB monoclonal antibody (1:400), mouse anti-IκB monoclonal antibody (1:400) and were purchased from Santa Cruz Biotechnology, Inc. The secondary antibodies used were horseradish peroxidase (HRP)-linked goat anti-rabbit immunoglobulin G (IgG) (1:4,000 dilution; Amersham Pharmacia Biotech, Piscataway, NJ, USA) and sheep anti-mouse IgG-HRP (1:8,000 dilution; Amersham Pharmacia Biotech). The blots were visualized by enhanced chemiluminescence (ECL) using a Pierce ECL western blotting substrate (Pierce Biotechnology, Inc.) and a Johnson enhanced chemiluminescence immunoassay analyzer (Shanghai Qian Jin Industrial Co., Ltd, Shanghai, China).