Rabbit anti phospho creb ser133

Cells were harvested in lysis buffer containing 120 mM NaCl, 0.5% Nonidet P-40, 0.2 mM sodium orthovanadate, 50 mM Tris–HCl pH 8.0, 1 Protease Inhibitor Cocktail Tablet per 50 ml (Roche Ltd, Basel, Switzerland). Extracts were centrifuged at 16,300 X g at 4°C for 15 min. Total protein content of samples was determined using the BioRad Protein Assay (BioRad Laboratories, Hercules, CA). Specific proteins were detected by Western blot analysis using methods described previously ^{(14 (link))} and the following antibodies: rabbit anti-ERK1/2 (Cell Signaling Technology, Danvers, MA), rabbit anti-phospho ERK1/2 (Cell Signaling Technology), rabbit anti-CREB (06-863 Millipore Billerica, MA), rabbit anti-phospho-CREB (Ser133) (06-519 Millipore Billerica, MA) rabbit anti-C/EBPβ (sc-150 Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-phospho-C/EBPβ (Thr235) (Cell Signaling Technology), mouse anti-β-actin (AC-74 Sigma-Aldrich St. Louis, MO), and mouse-anti-human VDR (sc-13133 Santa Cruz Biotechnology, Santa Cruz, CA). The membranes were incubated overnight with 1:1000 dilutions of the primary antibody prepared in blocking solution followed by a 1:5,000 dilution of horseradish-peroxidase-conjugated goat-anti-mouse IgG (Invitrogen, Carlsbad, CA) or mouse anti-rabbit IgG light chain specific secondary antibody (West Grove, PA).