Immunofluorescent analysis of CREB (green) in untreated HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 15 minutes at room temperature. Cells were then blocked with 5% normal goat serum (Product #31873) for 15 minutes at room temperature. Cells were probed with a mouse monoclonal antibody recognizing CREB (Product # MA1-083), at a dilution of 1:400 for at least 1 hour at room temperature. Cells were washed with PBS and incubated with DyLight 488 goat-anti-mouse secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei were stained with Hoechst 33342 dye (Product# 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Human, Mouse, Non-human primate, Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant human CREB-A protein.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.1-1.0 µg/mL|
|Immunoprecipitation (IP)||2 µg|
|Western Blot (WB)||1:500|
* 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.
MA1-083 detects the CREB in human, monkey, rat and mouse cells.
MA1-083 has been successfully used in immunofluorescence, Western blot, immunoprecipitation, immunohistochemistry, and ELISA procedures. By Western blot, this antibody detects a ~43 kDa protein.
The MA1-083 immunogen is human recombinant CREB-A protein.
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.