Rabbit anti tlr4 polyclonal antibody

Cells were seeded on coverslips and allowed to adhere for 12 h. After treatment, cells were fixed with 100 % methanol for 30 min and blocked with 5 % bovine serum albumin (BSA) for 30 min. Coverslips were incubated with mouse anti-HBx monoclonal antibody (1:100, abcam) or rabbit anti-TLR4 polyclonal antibody (1:100, Santa Cruz Biotechnology) in 1 % BSA at 4 °C overnight, followed by with fluorescence labeled secondary antibody (1:100, Earthox, USA) in 1 % BSA for 30 min at room temperature. Cell nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI; Sigma). Images were captured using Nikon A1 Confocal Laser Microscope System and adjusted using NIS-Elements Viewer 4.0 (Nikon, Japan).

Baicalin (pure, 98%) was purchased from Shanghai Xin Yu Biological Technology Co., Ltd (Shanghai, China). LPS (E. coli, serotype 055: B5) was purchased from Sigma (St. Louis, MO, USA). ELISA kits of IL-1β, TNF-α, and IL-6 were purchased from BioLegend (Camino Santa Fe, CA, USA), MPO and NO assay kits were obtained from the Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Rabbit anti-phospho-IkBα polyclonal antibody, rabbit anti-phospho-p65 polyclonal antibody and rabbit anti- p65 polyclonal antibody were purchased from Sangon Biotech Co., Ltd (Shanghai, China). Rabbit anti-TLR4 polyclonal antibody was obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Rabbit anti- β-actin polyclonal antibody was obtained from univ Biotechnology (Shanghai, China). Rabbit anti-lamin-B polyclonal antibody was obtained from Santa Cruz Biotechnology. And HRP-conjugated goat anti-rabbit IgG was obtained from ZSGE-BIO (Beijing, China). SYBR Green PCR Master Mix was purchased from Toyobo Life Science Co., Ltd (Shanghai, China). And M-MULV cDNA reverse transcription kit was purchased from Sangon Biotech Co., Ltd (Shanghai, China).