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Technical Data Sheet

CP4141

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CREB (N-terminal region)
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$245
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 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).

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Immunocytochemical labeling of CREB in A431 that were fixed in paraformaldehyde and permeabilized using NP-40. The cells were labeled with rabbit polyclonal CREB (N-terminal region). The antibody was detected using goat anti-rabbit DyLight® 594.

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Application
Dilution


ELISA
1:1000


ICC
1:100


IP
1:100


WB
1:500



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
CREB synthetic peptide (coupled to carrier protein) corresponding to amino acids in the N-terminal region of human CREB. This sequence is well conserved in rat, mouse, and chicken CREB, and has low homology to other CREB family members.
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, HeLa, and rat PC12 cells.

*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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