Analysis Note

ControlHeLa nuclear extract
Included Positive Antigen Control: Catalog # 12-309, HeLa nuclear extract. Add an equal volume of Laemmli reducing sample buffer to 5-10 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

Research Sub CategoryTranscription Factors

Research CategoryEpigenetics & Nuclear Function

Immunoprecipitation Analysis: 10 µg from a representative lot immunoprecipitated CREB in 500 µg of HeLa nuclear extract.
Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected an enhanced CREB occupancy at the LYPD3 promoter in LKB1-deficient TE-4 human esophageal cancer cells (Gu, Y., et al. (2012). Oncogene. 31(4):469–479).
Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected CREB association at the RNASET2 sites targeted by T-cell Leukemia Virus type 1 (HTLV-1) encoded Tax protein using HTLV-1-infected human T-cell lines, MT-2, C8166/45 and SLB-1 (Polakowski, N., et al. (2011). Viruses. 3(8):1485-500).
Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected CREB occupancy at the hBDNF promoter IV using postmortem human parietal cortex tissue chromatin preparations (Pruunsild, P., et al. (2011). J. Neurosci. 31(9):3295-3308).
Chromatin Immunoprecipitation (ChIP) Analysis: A representative lot detected CREB occupancy at the miR-9-2 promoter in SK-N-BE human neuroblastoma cells (Laneve, P., et al. (2010). Nucl. Acids Res. 38(20):6895-6905).
ChIP-seq Analysis: A representative lot identified CREB target genes in murine spermatogonia derived GC1-spg cells by ChIP-seq analysis (Martianov, I., et al. (2010). BMC Genomics. 11:530).
Western Blotting Analysis: Representative lots detected CREB in lysates from cultured embryonic rat striatal neurons and PC12 pheochromocytoma cells (Hutchinson, A.N., et al. (2012). Neuropsychopharmacology. 37(2):321-337; Maharjan, S., et al. (2010). J. Neurochem. 112(1):42-55).
Western Blotting Analysis: Representative lots detected CREB in mouse proximal caput epididymis-1 (PC-1) cell lysate and hippocampus tissue homogenates (Hamzeh, M., and Robaire, B. (2011). J. Endocrinol. 209(1):55-64; Zhou, Z., et al. (2009). Neuropharmacology. 56(1):81-89).
Western Blotting Analysis: A representative lot detected CREB in SK-N-BE human neuroblastoma cell lysate (Laneve, P., et al. (2010). Nucl. Acids Res. 38(20):6895-6905).
Electrophoretic Mobility Shift Assay (EMSA): A representative lot caused a supershift of DNA-CREB complex in EMSA using radiolabeled oligonucleotide containing hBDNF promoter I CREB-binding sequence and lysates from KCl-treated primary rat neurons (Pruunsild, P., et al. (2011). J. Neurosci. 31(9):3295-3308).

Anti-CREB Antibody, Cat. No. 06-863, is a highly specific rabbit Polyclonal antibody, that targets CREB and has been tested in Chromatin Immunoprecipitation (ChIP), ChIP-seq, Electrophoretic Mobility Shift Assay (EMSA), Immunoprecipitation, and Western Blotting.

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 cAMP-responsive element-binding protein 1, CREB-1) is encoded by the CREB1 gene (Gene ID 1385) in human. CREB is a transcription factor that binds the cAMP response elements (CRE) and regulates the transcription of the downstream genes. CREB activity is regulation via phosphorylation by various kinases, including PKA, and Ca2+/calmodulin-dependent protein kinases. Genes known to be regulated by CREB include c-fos, BDNF, tyrosine hydroxylase, neuropeptides (such as somatostatin, enkephalin, VGF, and corticotropin-releasing hormone), as well as the circadian clock genes PER1 and PER2. CREB is critical for memory formation across a wide range of species.

Immunogen

Epitope: Amino acids 5-24.

KLH-conjugated, synthetic peptide (CSGAENQ QSGDAAVTEAENQQ) corresponding to a.a. 5-24 of human CREB. The immunizing sequence is identical in pig, shares 19 of 20 residues with mouse and bovine and 18 of 20 residues with rat.

Legal Information

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

Linkage

Replaces: 04-218

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein A purified

Purified rabbit polyclonal antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide with 30% glycerol.

Quality

Evaluated by Western Blotting in HeLa nuclear extract.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected CREB in 10 µg of HeLa nuclear extract.

Specificity

This antibody recognizes CREB.

Storage and Stability

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.

Target description

~43 kDa observed.