Catalog # CK6460

CREB Phospho-Regulation Antibody Kit

Antibody Sampler Kit



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.

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.

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 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.

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.

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.

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

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

This kit contains:

CREB (N-terminal region) Rabbit pAb50 μlWB, E, IP, ICCHu, Rt, Ms, Ck45
CREB (Ser-133), phospho-specific Mouse mAb50 μlWB, E, ICCHu45
CREB (Ser-271), phospho-specific Rabbit pAb 50 μlWB, E, ICCHu, Rt, Ms, Ck45
Anti-Mouse Ig:HRPDonkey pAb100 μlWB, EMs
Anti-Rabbit Ig Light ChainSpecific:HRP Mouse mAb100 μlWB, E, ICC, IHCRb

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.