Immunofluorescent analysis of CREB (green) showing staining in the nucleus of SK-N-MC cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CREB polyclonal antibody (Product # PA1-850) in 3% BSA-PBS at a dilution of 1:200 and incubated overnight at 4°C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. Actin was stained using Alexa Fluor 554 (red) and nuclei were stained with Hoechst or DAPI (blue). Images were taken at a magnification of 60x.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Rat, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues K(123) R R E I L S R R P S Y R K(136) of human CREB.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||1-3 ul|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Western Blot (WB)||2 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.
|Western Blot (WB)||See 2 publications below|
PA1-850 detects both the phosphorylated and non-phosphorylated forms of cyclic-AMP response element binding protein (CREB) from rat, mouse, and human samples.
PA1-850 has been successfully used in ChIP, Western blot, ICC/IF and IHC-P procedures. By Western blot, this antibody detects an ~43 kDa protein representing CREB from GH4 cell extract.
The PA1-850 immunogen is a synthetic peptide corresponding to residues K(123) R R E I L S R R P S Y R K(136) of human CREB. This sequence is completely conserved between human, bovine, canine, mouse, pig, sheep, and rat CREB protein. The PA1-850 immunizing peptide (Cat. # PEP-142) is available for use in neutralization and control experiments.
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.
Distinct cAMP response element-binding protein (CREB) domains stimulate different steps in a concerted mechanism of transcription activation.
PA1-850 was used in western blot to identify the functional roles of CREB different domains in transcriptional activation
|Kim J,Lu J,Quinn PG||Proceedings of the National Academy of Sciences of the United States of America (97:11292)||2000|
Defective thymocyte proliferation and IL-2 production in transgenic mice expressing a dominant-negative form of CREB.
PA1-850 was used in western blot to investigate the functional consequences of a dominant-negative form of CREB
|Barton K,Muthusamy N,Chanyangam M,Fischer C,Clendenin C,Leiden JM||Nature (379:81)||1996|