Rabbit anti il8

Virus induced supernatants from infected cells were treated with rabbit anti-IL-6 or rabbit anti-IL8 or (normal rabbit serum) control antibodies (all from Abcam) at concentrations specified for 1 hr before treatment of cell monolayers. Treated supernatents were left in place throughout the experiment.

The dewaxed slide was treated with Proteinase K (Sigma-Aldrich) for proteolytic digestion and 3% H₂O₂ for the elimination of endogenous peroxidase activity. After blocking with normal goat serum, the slides were incubated with primary antibody overnight at 4 °C, respectively. The primary antibodies used in this study include: rabbit anti-OCN (Abcam), rabbit anti-IL-8 (Abcam), rabbit anti-CCL5 (Abcam), rabbit anti-IL-1β (Abcam), rabbit anti-IL-1ra (Abcam), anti-CD68 (Abcam), rabbit anti-Phospho-Histone H3S10 (Abcam), and rabbit anti-TRPM7 (Abcam). The slides were then incubated with goat anti-rabbit secondary antibody and visualized using Diaminobenzidine (DAB) staining kit (Santa Cruz Biotechnology, Santa Cruz, USA) following the manufacturer’s instruction. Immunofluorescent staining was done using Alexa-Fluor 488 conjugated anti-rabbit IgG or Alexa-Fluor 647 conjugated anti-mouse IgG secondary antibodies (Thermo Fisher Scientific) and Hoechst 33324 (Thermo Fisher Scientific). Immunofluorescent images were captured using an LSM 780 confocal microscopy (Zeiss, Germany)