Anti nlrc4

Immunofluorescence assay was performed as described in research by Hou et al [18 (link)]. Sections were permeabilized with Triton X-100 (0.1%) and blocked with solution containing 5% bovine serum before applying primary antibody. Specimens were incubated respectively with anti-NLRC4 (Invitrogen, cat #PA5-88997; 1:100) and anti-caspase-1 (Invitrogen, cat #MA5-32909; 1:100) for 12 hours in 4°C. Secondary antibodies (Life Technologies, catalog #A21207; 1:200) and Alexa Fluor 488 goat anti-mouse IgG (LifeTechnologies, catalog #A11001) were incubated subsequently under light-protected conditions for 1 h at room temperature. Nucleus were staining with DAPI (KeyGen Biotech, catalog #KGA215–10) in the end. After final washing, the cover slips were mounted on slides using 50% glycerin. Then the sections were observed under a fluorescence microscope (Olympus) or confocal laser scanning microscope (Olympus).

For IHC, after antigen retrieval using EDTA, the specimens were blocked with goat serum for 20 minutes before applying the primary antibody. Specimens were incubated with anti-NLRC4 (Invitrogen, cat #PA5-88997; 1:100) for 12 hours in 4°C. Next, the sections were washed twice and subsequently incubated with HRP-polymer-conjugated secondary antibody (Zhong Shan, China) at room temperature. Finally, the specimens were then stained with 3,3-diaminobenzidine solution and haematoxylin. The slides were photographed with an inverted microscope (Olympus, Japan).