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Technical Data Sheet

CP4161

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CREB (Ser-271), phospho-specific
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms, Ck
43 kDa

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Western blot analysis of human A431 cells treated with calyculin A (100 nM) for 30 min. (lanes 1, 3, & 5) then 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).

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Immunocytochemical labeling of phosphorylated CREB in control and calyculin A-treated A431 cells. The cells were fixed in paraformaldehyde and permeabilized using NP-40 before labeling with rabbit polyclonal CREB (Ser-271). The antibody was detected using goat anti-rabbit DyLight® 594.

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Application
Dilution


ELISA
1:2000


ICC
1:200


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
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.


Background References
Gonzalez, G.A. & Montminy, M.R. (1989) Cell. 59(4):675.
Lonze, B.E. & Ginty, D.D. (2002) Neuron 35:605.
Sakamoto, K. et al. (2010) Mol. Biol. Cell 21:2966.
Immunogen
Phospho-CREB (Ser-271) synthetic peptide (coupled to carrier protein) corresponding to amino acids surrounding Ser-271 in human CREB. This sequence is well conserved in rat, mouse, chicken, and zebrafish CREB. The site has some homology to the conserved site in ATF1 (Ser-198) and CREM (Ser-287).
Buffer and Storage
Rabbit polyclonal, affinity-purified antibody 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.
Specificity
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, but 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.
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