Analysis Note

ControlForskolin-treated PC12 cell lysate

Application

Research Sub CategoryTranscription Factors

ELISA: Recommended at 0.05 µg/mL by an independent laboratory.

Research CategoryEpigenetics & Nuclear Function

Anti-phospho-CREB (Ser133) Antibody, clone 10E9 is a Mouse Monoclonal Antibody for detection of phospho-CREB (Ser133) also known as active transcription factor CREB & has been validated in WB, ELISA.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Cyclic AMP-responsive element-binding protein 1 (UniProt: P16220; also known as CREB-1, cAMP-responsive element-binding protein 1) is encoded by the CREB1 gene (Gene ID: 1385) in human. CREB is a phosphorylation-dependent transcription factor that stimulates transcription upon binding to the DNA cAMP response element (CRE; TGACGTCA) as a dimer. It is essential for the activation of a number of immediate early genes. It also plays a key role in promoting neuronal survival, differentiation, proliferation, and neurite outgrowth. Transcription activation is reported to be enhanced by the TORC coactivators. CREB has four main functional domains: a C-terminal basic/leucine zipper domain (bZIP; aa 269-327), the Kinase Inducible Domain at the N-terminal (KID; aa 87-146), and two glutamine-rich domains (Q1, and Q2). The Q1 and Q2 domains provide transactivation function in the absence of cAMP signaling and synergize with KID and bZIP to confer full activity to CREB in response to cAMP. CREB is phosphorylated on serine 133 by protein kinase A, which promotes the recruitment of the co-activator CREB-binding protein(CBP) and p300 that increases the transcription of CREB-dependent genes. CREB is also phosphorylated on serine 133 by mitogen- and stress-activated kinase 1/2 (MASK 1/2) in cells in response to the activation of MAPK signaling. Phosphorylation of CREB is sufficient to induce cellular gene expression in response to cAMP, however, additional promoter-bound factors are required for target gene activation by CREB in response to mitogen or stress signals, indicating the presence of components in the CREB pathway that discriminate between different types of inputs. (Ref.: Wang, H., et al. (2018). Front. Mol. Neurosci. 11; 255; Naqvi, S., et al. (2014) Biochem. J. 458(3); 469-479; Mayr, B., and Montiminy, M. (2001). Nat. Rev. Mol. Cell Biol. 2(8); 599-609)..

Immunogen

KLH-conjugated, synthetic peptide (REILSRRP(pS)YR) corresponding to amino acids 125-135 of human CREB. The immunizing sequence is identical in human and mouse. Clone 10E9.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Physical form

Thiophilic adsoption and size exclusion chromatography

Purified mouse monoclonal IgG1κ from serum-free cell culture supernatant, lyophilized from 1 mL of PBS, 0.09% sodium azide, PEG, and sucrose. Lyophilized.

Format: Purified

Quality

Evaluated by western blot on whole cells lysates from rat PC12 cells treated with forskolin.

Western Blot Analysis: 0.5-2 µg/mL of this antibody detected phosphorylated CREB in whole cell lysates from HepG2 and PC12 cells treated with forskolin.

Specificity

Recognizes phosphorylated CREB at Mr 46 kDa. May also recognize p30 and p38 proteins, which likely correspond to phosphorylated CREM and ATF-1 respectively, since the two proteins share significant homology with the phosphorylated CREB immunizing peptide

Storage and Stability

Lyophilized: Stable for 1 year at -20°C from date of receipt.
Rehydrated: Stable for 3 months at 2-8°C.
Aliquot solution to store frozen and avoid repeated freeze thaw cycles.

Target description

46 kDa