15 μg of U-2 OS lysate was run on 6-18% SDS-PAGE under reducing conditions and blotted onto nitrocellulose membrane. antibody at 1 μg/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. IL18 band was visualized using ECL Substrate.

Result: antibody can detect IL18 by Western blotting.

IHC-P analysis of tonsil tissue by IL18 antibody.
IHC-P was performed using sections of the formalin-fixed paraffin-embedded tonsil tissue. Antigen was retrieved through addition of boiling Tris/EDTA buffer pH 9 in a pressure cooker for 3 min. Endogenous peroxidase activity was quenched by incubating the sections with 3% H₂O₂ for 30 min at room temperature. The sections were then incubated with IL18 antibody at 2 μg/mL at room temperature for 1 h. Poly-peroxidase conjugated goat anti-mouse IgG was used as the secondary antibody. Diaminobenzidine was used as the chromogen. The section was counterstained with hematoxylin. A tissue section incubated with phosphate-buffered saline followed by incubation with the secondary antibody was used as the background control.

Result: Germinal center cells, non-germinal center cells and squamous epithelial cells are positively stained at the cytoplasm.

Immunoprecipitation was performed by incubation of 2.5 μg antibody with U-2 OS lysate containing 200 μg total protein. After absorption with Protein G beads, the mixture was run on 6-18% SDS-PAGE and blotted onto nitrocellulose membrane. IL18 at 1 μg/mL was used as the primary antibody and peroxidase conjugated goat anti-mouse IgG (Fc specific) was used as the secondary antibody.

Lane 1: U-2 OS lysate

Lane 2: IL18 immunoprecipitated from U-2 OS lysate by antibody

Lane 3: The same as Lane 2 but KT82 was used as IgG isotype control antibody

Result: antibody can immunoprecipitate IL18.