Catalog # CM4151

CREB (Ser-133), phospho-specific Antibody

Mouse Monoclonal

Application / Dilution

ELISA	1:2000
ICC	1:100
WB	1:1000

Size 100 μl

Species Reactivity Hu, Rt, Ms, Ck

MW 43 kDa

Isotype IgG1

CREB (cyclic AMP response element-binding protein) is a stimulus-induced transcription factor that plays pivotal roles in cell survival and proliferation. CREB is expressed in various tissues, and has important gene-regulating roles in the nervous system. The transactivation function of CREB is primarily regulated through Ser-133 phosphorylation by cAMP-dependent protein kinase A (PKA) and related kinases. CREB is phosphorylated at other sites in response to calcium influx and DNA damage. The DNA-damage responsive nuclear kinase, HIPK2, can phosphorylate Ser-271 but not Ser-133 in CREB, and this phosphorylation activates CREB transactivation function. Mutation of Ser-271 to Glu-271 potentiates the CREB transactivation function. Thus, phosphorylation of Ser-271 may be the mode of activation for CREB-dependent transcription in response to genotoxic stress.

References

Sakamoto, K. et al. (2010) Mol. Biol. Cell 21:2966.
Lonze, B.E. & Ginty, D.D. (2002) Neuron 35:605.
Gonzalez, G.A. & Montminy, M.R. (1989) Cell. 59(4):675.

Western blot analysis of human A431 cells treated with calyculin A (100 nM) for 30 min. (lanes 1, 3 & 5) then the blots were treated with lambda phosphatase (lanes 2, 4 & 6). The blots were probed with anti-CREB (N-terminal region) (lanes 1 & 2), anti-CREB (Ser-133) (lanes 3 & 4), and anti-CREB (Ser-271) (lanes 5 & 6).

Immunocytochemical labeling of phosphorylated CREB in calyculin A-treated A431 cells. The cells were fixed in paraformaldehyde and permeabilized using NP-40 before labeling with mouse monoclonal CREB (Ser-133). The antibody was detected using goat anti-mouse DyLight® 594.

Clone M415 was generated from phospho-CREB (Ser-133) synthetic peptide (coupled to carrier protein) corresponding to amino acids surrounding Ser-133 in human CREB. This sequence is well conserved in rat, mouse, chicken, and zebrafish CREB. The site is also conserved in CREM (Ser-136) and ATF1 (Ser-63).

*For more information, see UniProt Accession P16220

Mouse monoclonal antibody purified with protein A chromatography is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

The antibody detects a 43 kDa* protein corresponding to the molecular mass of CREB on SDS-PAGE immunoblots of human A431, Jurkat, and rat PC12 cells treated with Calyculin A, as well as A431 treated with EGF and PC12 treated with NGF or Forskolin. This reactivity is not observed after lambda phosphatase treatment.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains: