Mmp 2 antibody

Mice aorta were collected and fixed with formalin. After paraffin embedding, the tissues were and cut into 5 μm sections slides. Then, the slides were deparaffinized and rehydrated using xylene and gradient ethanol, treated with Tris/EDTA pH 9.0 buffer for heat-induced epitope retrieval. Next, the slides were treated with 3% hydrogen peroxide to quench endogenous peroxidase and blocked with 10% goat serum to reduce nonspecific binding. Followingly, section slides were incubated with MMP-2 antibody (Proteintech, Cat#10373-2-AP) and then HRP-linked anti-rabbit IgG (1:10,000, Cat#7074S, Cell Signaling Technology). After washing with tris-buffered saline tween-20 (TBST), the slides were incubated with 3,3′-diaminobenzidine (DAB) (Beyotime, Cat#P0203) and observed under a fluorescence microscope (Olympus IX53, Tokyo, Japan).

Paraformaldehyde-fixed paraffin-embedded sections were incubated with primary antibodies overnight at 4°C. The primary antibodies used were against OPN-N (rabbit anti-OPN-N antibody, ab181440, 1:200; Abcam, United Kingdom), NLRP3 (rabbit anti-NLRP3 antibody, 1:200; Boster, Wuhan, China), ASC (rabbit anti-ASC antibody, 10500-1-AP, 1:200; Proteintech, Wuhan, China), IL-18 (rabbit anti- IL-18 antibody, 1:200; Proteintech), IL-1β (rabbit anti-IL-1β antibody, 1:200; Arigo, Hamburg, Germany), Caspase-1 (rabbit anti- Caspase-1 antibody, 22915-1-AP, 1:200; Proteintech), MMP2 (rabbit anti-MMP2 antibody, 1:200; Proteintech), MMP9 (rabbit anti-MMP9 antibody, 1:200; Proteintech), α-smooth muscle actin (SMA) (rabbit anti-α-SMA, 1:200; Proteintech) and α9β1 (rabbit anti-α9β1, ab27947, 1:200; Abcam). As negative controls, species and isotype-matched IgG was applied in place of the primary antibody. Immunohistochemical analysis was performed using the SP-9001 SPlink Detection kits (OriGene Technologies, Inc.), according to the manufacturers’ instructions. All sections were examined using a Nikon E100 microscope. Three sections were randomly selected from each group.