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Technical Data Sheet

CK6460

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CREB Phospho-Regulation
Antibody Sampler Kit
Price

$395

Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
Kit Summary
The CREB phospho-regulation antibody sampler kit can be used to examine Ser-133 and Ser-271 phosphorylation in CREB. The kit also includes a rabbit polyclonal antibody to monitor total expression levels of CREB, and HRP-conjugated secondary reagents.
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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 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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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.
Buffer and Storage
Mouse monoclonal and rabbit polyclonal antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.
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