Mouse anti s6 antibody

Immunoblotting was performed as described previously.^{31 (link)} In brief, whole-cell lysates were prepared using boiling lysis buffer (1% SDS, 10 mM Tris·HCl, pH 7.4). Equal amounts of proteins were separated using Criterion or mini gradient polyacrylamide gels (Bio-Rad, Hercules, CA, USA) and transferred to PVDF membranes. The following rabbit antibodies for: phospho-S6 (S235/236), phospho-S6 (S240/244), phospho ERK½, phospho-Thr 389 p70S6K, phospho-AKT (S473) and phospho-AKT (T308), phospho-IRS1 (S1101), phospho-IRS1(S636/639), IRS1 and mouse anti-S6 antibody were from Cell Signaling Biotechnology (Danvers, MA, USA). Rabbit anti-actin and mouse monoclonal anti-GAPDH antibodies were from Sigma-Aldrich and Invitrogen (Camarillo, CA, USA), respectively. Secondary anti-rabbit and anti-mouse HRP-conjugated antibodies were from Cell Signaling Biotechnology.

Western blot analysis was performed under conventional conditions. The antibodies used were as follows: a rabbit anti-human S100A11 antibody that we made17–19 (link), mouse anti-human S100A11 antibody (MBL, Nagoya, Japan), rabbit anti-S100A6 antibody (Cell Signaling Technology, Beverly, MA, USA), rabbit anti-calgranulin A (S100A8) antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA), rabbit anti-calgranulin B (S100A9) antibody (Santa Cruz Biotechnology), mouse anti-RAGE antibody (R&D Systems), mouse anti-tubulin antibody (Sigma-Aldrich, St. Louis, MO, USA), mouse anti-fibroblast antibody (EMD Millipore, Temecula, CA, USA), mouse anti-α-smooth muscle actin antibody (DAKO Agilent Technologies, Santa Clara, CA, USA), rabbit anti-phospho-p70 S6 kinase antibody (Cell Signaling Technology), rabbit anti-p70 S6 kinase antibody (Cell Signaling Technology), rabbit anti-phospho-S6 antibody (Cell Signaling Technology), mouse anti-S6 antibody (Cell Signaling Technology), mouse anti-HA antibody (Cell Signaling Technology), and goat anti-GST antibody (GE Healthcare Bio-Sciences, Piscataway, NJ, USA).