Cell Signaling Technology

Product Pathways - Neuroscience

CREB (D76D11) XP™ Rabbit mAb #4820

This XP™ monoclonal antibody was developed using our eXceptional Monoclonal Technology (XMT™).

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC F ChIP H M R Mk Hm Dm Endogenous 43 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Hm=Hamster  Dm=D. melanogaster
Species cross-reactivity is determined by Western blot.

Protocols

Specificity / Sensitivity

CREB (D76D11) XP™ Rabbit mAb detects endogenous levels of total CREB protein. It cross reacts with ATF-1

Source / Purification

Monoclonal antibody is produced by immunizing animals with a full length GST-CREB fusion protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using CREB (D76D11) XP™ Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of SH-SY5Y cells, using CREB (D76D11) XP™ Rabbit mAb Antibody (blue) compared to concentration matched Rabbit (DA1E) mAb IgG XP™ Isotype Control #3900 (red).

IF-IC

IF-IC

Confocal immunofluorescent analysis of SH-SY5Y cells using CREB (D76D11) XP™ Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293 cells treated with Forskolin #3828 (30 µM) and either 10 μl of CREB (D76D11) XP™ Rabbit mAb #4820 or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP™ Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by Real-Time PCR using primers specific for the ALS2 and NR4A3 genes, and the heterochromatic α Satellite repeat element. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Background

CREB is a bZIP transcription factor that activates target genes through cAMP response elements. CREB is able to mediate signals from numerous physiological stimuli, resulting in regulation of a broad array of cellular responses. While CREB is expressed in numerous tissues, it plays a large regulatory role in the nervous system. CREB is believed to play a key role in promoting neuronal survival, precursor proliferation, neurite outgrowth and neuronal differentiation in certain neuronal populations (1-3). Additionally, CREB signaling is involved in learning and memory in several organisms (4-6). CREB is able to selectively activate numerous downstream genes through interactions with different dimerization partners. CREB is activated by phosphorylation at Ser133 by various signaling pathways including Erk, Ca2+ and stress signaling. Some of the kinases involved in phosphorylating CREB at Ser133 are p90RSK, MSK, CaMKIV and MAPKAPK-2 (7-9).

  1. Lonze, B.E. et al. (2002) Neuron 34, 371-85.
  2. Lee, M.M. et al. (1999) J Neurosci Res 55, 702-12.
  3. Redmond, L. et al. (2002) Neuron 34, 999-1010.
  4. Dash, P.K. et al. (1990) Nature 345, 718-21.
  5. Yin, J.C. et al. (1994) Cell 79, 49-58.
  6. Guzowski, J.F. and McGaugh, J.L. (1997) Proc Natl Acad Sci USA 94, 2693-8.
  7. Xing, J. et al. (1998) Mol Cell Biol 18, 1946-55.
  8. Ribar, T.J. et al. (2000) J Neurosci 20, RC107.
  9. Tan, Y. et al. (1996) EMBO J 15, 4629-42.

Application References

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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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