ECM Logo Bar
Search :
Technical Data Sheet

CM4151

image
image
image
CREB (Ser-133), phospho-specific
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$245
100 μl
Hu, Rt, Ms, Ck
43 kDa

image
image
Enlarge

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).

image
Enlarge

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.

image
Application
Dilution


ELISA
1:2000


ICC
1:100


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.
image
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
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).
Buffer and Storage
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.
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, 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.
imageRelated Products
CK6460
CREB Phospho-Regulation Antibody Sampler Kit

CP4141
CREB (N-terminal region) Rabbit Polyclonal

CP4161
CREB (Ser-271), phospho-specific Rabbit Polyclonal

CX4165
phospho-CREB (Ser-271) Blocking Peptide

MK6050
MAP Kinase Activation Antibody Sampler Kit

PK6140
p38 MAPK Phospho-Regulation Antibody Sampler Kit

New Releases
AM0171
image
Annexin A5
Mouse Monoclonal
image
CK6740
image
VE-Cadherin Phospho-Regulation
Antibody Sampler Kit
image
MM0271
image
Mitofilin
Mouse Monoclonal
image