Anti il 18 10663 1 ap

Frozen slices were fixed with 4% paraformaldehyde for 15 min and incubated with 0.3% Triton X-100 for 15 min. The slices were incubated with 3% hydrogen peroxide for 10 min, and nonspecific epitopes were blocked with goat serum for 15 min. Each slice was incubated with the following primary antibodies: anti-IL-1β (A11370; Abclonal), anti-IL-18 (10663-1-AP; Proteintech), anti-VCAM-1 (11444-1-AP; Proteintech), anti-ICAM-1 (10020-1-AP; Proteintech), anti-IL-6 (66146-1-1g; Proteintech), anti-TNF-α (A11534; Abclonal), and anti-CD68 (25747-1-AP; Proteintech). After overnight incubation, the slices were incubated with Streptomyces anti-biotin peroxidase solution for 1 h at 37 ?. The slices were then visualized using DAB solution (Zhongshanjingqiao Biotechnical) and counterstained with hematoxylin for 8 min. Finally, all the slides were examined under a light microscope, and images were analyzed using Image Pro Plus software.

Cell lysates were incubated with IgG or anti-SCAP antibody for 2?h at 4?°C, and then magnetic beads were bound to the SCAP immune complexes at 4°C overnight. After incubation, the magnetic beads were washed with phosphate-buffered saline; the proteins were collected from the magnetic beads, mixed with protein loading buffer, and boiled for 8 min at 100°C. Finally, the proteins were detected by Western blotting analysis.
The proteins were separated by SDS-PAGE and then transferred to a PVDF membrane. After that, the membranes were blocked with 3% bovine serum albumin for 2 h, followed by incubation with the following primary antibodies: anti-nSREBP2 (ab30682; Abcam), anti-caspase 1 (22915-1-AP; Proteintech), anti-cleaved caspase-1 (4199S, Cell Signaling Technology), anti-IL-1β (A11370; Abclonal), anti-IL-18 (10663-1-AP; Proteintech), anti-VCAM-1 (11444-1-AP; Proteintech), anti-ICAM-1 (10020-1-AP; Proteintech) and anti-NLRP3 (NBP1-97601; Novus Biologicals). After overnight incubation at 4°C, the membranes were incubated with horseradish peroxidase-labeled goat anti-rabbit with rotation for 2 h at 37°C. Finally, detection was performed using an ECL chemical luminescent detection kit (Bio-Rad), and the bands were further analyzed using Quantity One software. The expression of the target protein was normalized to β-actin expression.