Rabbit anti cox 2 antibody

Tissues and A549 cells were harvested, lysed in RIPA buffer (Solarbio, Beijing, China) containing protease inhibitor PMSF (Solarbio). Protein concentrations were determined using a BCA kit (Thermo Fisher Scientific, USA). Proteins were separated on 8% or 12% SDS-PAGE Gels. Separated proteins were transferred onto polyvinylidene difluoride (PDVF) membranes, which were blocked with 5% non-fat milk in TBST and incubated with the primary antibodies overnight at 4°C. Subsequently, membranes were incubated with appropriate secondary HRP-linked antibodies. Proteins were visualized by enhanced chemiluminescence (Millipore, Burlington, MA, USA). Images were obtained using ChemiDoc XRS (Bio-Rad). The relative band intensity was quantified using the Image Lab Analyzer software (Bio-Rad, Hercules, CA). The antibodies used in the present research were as follows: rabbit anti-β-Tubulin antibody and rabbit anti-GAPDH antibody (1:2000, Servicebio, Wuhan, China); rabbit anti-α-SMA antibody (1:1000, SAB, Maryland, USA); rabbit anti-sEH antibody (1:5000, Abcam, USA); rabbit anti-COX-2 antibody (1:1000, Servicebio); rabbit anti-Collagen Type I antibody (1:1000, Proteintech, Rosemont, USA); rabbit anti-Collagen Type III antibody (1:1000, Proteintech); rabbit anti-SFTPC antibody (1:1000, Abcam); rabbit anti-p53 antibody (1:3000, Proteintech).

As described in our previous study [24 (link)]. Frozen mouse liver tissue was taken out and lysed in cold RIPA buffer (Solarbio, Beijing, China) containing a protease inhibitor (Roche, Mannheim, Germany). Protein quantification was performed using a bicinchoninic acid (BCA) protein assay kit (Thermo Fisher Scientific), and the protein (30 mg) in each sample was separated by 12% SDS-PAGE, and then transferred to 0.45-μm polyvinylidene difluoride (PVDF) membranes. The membranes were blocked with 5% fat-free milk and incubated with the primary antibodies overnight at 4°C. Subsequently, membranes were incubated with appropriate secondary HRP-linked antibodies. The proteins were detected with a gel imaging system (Bio-Rad). The antibodies used in present research were as follows: rabbit anti-COX-2 antibody (1:1000, Servicebio, Wuhan, China), rabbit anti-sEH antibody (1:5000, Abcam, USA), rabbit anti-β-tubulin antibody (1:2000, Servicebio), rabbit anti-α-SMA antibody (1:1000, SAB, Maryland, USA), rabbit anti-NLRP3 antibody (1:2000, CST, USA), rabbit anti-pro-caspase-1 antibody (1:1000, Abcam). The expression of protein was normalized to β-tubulin.