|Detection of Human Phospho-CREB (S133) by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and HEK293 human embryonic kidney cell line untreated (-) or treated (+) with 20 mJ/cm2 ultraviolet light (UV) with a 30 minute recovery. PVDF membrane was probed with 0.5 µg/mL of Mouse Anti-Human Phospho-CREB (S133) Monoclonal Antibody (Catalog # MAB6906) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-CREB (S133) at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|Phospho-CREB (S133) in HeLa Human Cell Line. CREB phosphorylated at A133 was detected in immersion fixed Hela human cervical epithelial carcinoma cells, unstimulated (lower panel) and stimulated (upper panel) with PMA, using Mouse Anti-Human Phospho-CREB (S133) Monoclonal Antibody (Catalog # MAB6906) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
The cAMP response element binding protein (CREB) belongs to the bZIP superfamily of transcription factors, containing a basic domain that mediates DNA binding and a leucine zipper domain that facilitates dimerization. Within the promoter of target genes, CREB dimers bind cAMP response elements, defined by the palindromic consensus sequence TGACGTCA. When phosphorylated at Ser133, CREB also binds the coactivator CREB binding protein (CBP), which enhances transcription by acetylating histones to facilitate chromatin unraveling.