Anti il 23p19 antibody

Frozen tissue samples were fixed in 3.7% formaldehyde (FA) overnight at 4°C, transferred into 1% FA until they were embedded in paraffin. Paraffin samples were cut into 5 µm sections and stained with hematoxylin and eosin. Tumor grades in histological sections were classified into aberrant crypt foci (ACF), adenoma (Ad), and mucosal carcinoma. Detection of IL-23 in the tumor tissue was performed with an anti-IL-23p19 antibody (rabbit polyclonal, Abcam). Proliferating cells were detected via Ki-67 staining with a rat monoclonal antibody (Dako). Secondary anti-rabbit and anti-rat (Vector Laboratories, Inc.) antibodies coupled to biotin were added and sections were developed via the streptavidin–HRP VECTASTAIN^® ABC Kit (Vector Laboratories, Inc.). Finally, color development was performed with 3-amino-9-ethylcarbazole (AEC; Dako). All sections were counterstained with Harris hematoxylin solution (Sigma Aldrich).

IMQ cream (Beselna cream) was kindly provided by Mochida Pharmaceutical (Tokyo, Japan). Recombinant human TNF-α, IL-17A, and leptin were purchased from Chemicon (Temecula, CA, USA), R&D Systems (Minneapolis, MN, USA), and PeproTech (Cranbury, NJ, USA), respectively. The BSA–palmitate saturated fatty acid complex and BSA control were purchased from Cayman Chemical (Ann Arbor, MI, USA). Anti-STAT3 (clone: D1B2J, #30835), anti-phospho-STAT3 (Tyr705, clone: D3A7, #9145), anti-phospho-JNK (clone: 81E11, #4668), anti-total-JNK (#9252), anti-phospho-p38 (clone: D3F9, #4511), anti-total-p38 (clone: D13E1, #8690), anti-phospho-NF-κB p65 (clone: 93H1, #8214), and anti-total-NF-κB p65 (clone: D14E12, #8242) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-IL-23p19 antibody (#ab45420) was obtained from Abcam (Waltham, MA, USA). Anti-actin antibody was purchased from Sigma-Aldrich (A2066, St. Louis, MO, USA).