Immunofluorescent analysis of Phospho-CREB pSer133 (green) in HeLa cells either left untreated (left panel) or treated with 1uM Camptothecin (right panel) for 20 hours. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a Phospho-CREB pSer133 monoclonal antibody (Product # MA1-114) at a concentration of 4ug/ml for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan and ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Dog, Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||A synthetic phosphopeptide corresponding to amino acids 125-135 surrounding the Ser133 phosphorylation site of human CREB|
|Storage buffer||PBS, pH 7.2, with sucrose, PEG, 50% glycerol|
|Contains||0.09% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.05 µg/ml|
|Flow Cytometry (Flow)||1 ug/test|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50-1:500|
|Western Blot (WB)||2-4 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody recognizes CREB protein phosphorylated at serine 133 and ATF1 phosphorylated at serine 63. The antibody was purified from serum-free cell culture supernatant by subsequent thiophilic adsorption and size exclusion chromatography.
Aliquot and freeze in liquid nitrogen. Antibody can be stored frozen at -80°C for up to 1 year. Thaw aliquots at 37°C; thawed aliquots may be stored at 4°C for up to 3 months. Avoid repeated freeze/thaw cycles.
This antibody was originally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.
Cyclic AMP response element binding protein (CREB) is a basic/leucine zipper transcription factor that binds the cyclic AMP response element (CRE) and activates transcription in response to a variety of extracellular signals including neurotransmitters, hormones, membrane depolarization, and growth and neurotrophic factors. Activation of CREB is dependent upon the phosphorylation of serine 133. Phosphorylation occurs via p44/42 MAP kinase and p90RSK and also via p38 MAP kinase and MSK1. Although CREB will bind DNA independent of its phosphorylation state, only the phosphorylated form is competent as a transcription factor. CREB binding protein (CBP), a transcriptional coactivator that directly interacts with CREB, binds to CREB in the region of serine 133. CREB appears to play an important role in learning and memory.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.