Rabbit anti phosphorylated stat3 tyr705

Fetal lungs at e14.5, e16.5 and e18.5 (6 on each day) were isolated, fixed in phosphate-buffered 4% paraformaldehyde solution and embedded in paraffin. Paraffin-embedded sections were rehydrated through xylene and graded ethanol solutions. After treatment with 0.3% hydrogen peroxide, the sections were heated at 95°C for 40 min in antigen retrieval solution (pH 8; Nichirei, Tokyo, Japan). After treatment with 4% Blockace in PBS (Dai-Nippon-Pharmaceutical, Suita, Japan) for 20 min, the sections were treated with goat anti-Clara cell secretory protein (CCSP; Santa Cruz Biotech, Santa Cruz, CA), mouse anti-Foxj1 (Santa Cruz), goat anti-calcitonin gene-related peptide (CGRP; Abcam, Cambridge, MA), rabbit anti-Hes1 (Santa Cruz) or rabbit anti-phosphorylated Stat3 (Tyr705) (Cell Signaling) antibody overnight at 4°C. After washing, the sections were treated with anti-goat, -rabbit or -mouse IgG conjugated with HRP-polymer (ImmPress Reagent; Vector Laboratories, Burlingame, CA) for 30 min at room temperature. The sections were further treated with diaminobenzidine-hydrogen peroxide solution, and counterstained with hematoxylin.