Rabbit anti phospho creb ser133 antibody

Chromatin immunoprecipitation (ChIP) assays were performed using a SimpleChIP Enzymatic Chromatin IP Kit (9003, Cell Signaling Technologies). BMDMs from WT mice treated with 200 ng/ml LPS for 1 h were cross linked with 37% formaldehyde at a final concentration of 1% at room temperature for 10 min and glycine solution was added to stop the cross-linking reaction. Fragmented chromatin was treated with nuclease and subjected to sonication. Chromatin immunoprecipitation was performed overnight at 4 °C with rabbit anti- Phospho-CREB (Ser133) antibody (1:50, 9198, Cell Signaling Technologies) and normal rabbit IgG (1:250, 2729, Cell Signaling Technologies) as a negative control. Protein G Magnetic Beads were added for another 2 h at 4 °C. The chromatins were then washed and eluted from the protein G magnetic beads using buffers supplied with the kit and DNA was analyzed by quantitative PCR. The primers are listed in Supplemental Table 3 (Table S3).