Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-CREB (Ser133) (87G3) Rabbit mAb (Alexa Fluor® 488 Conjugate) #9187

Applications Reactivity Source Isotype
IF-F IF-IC F H M Rabbit IgG

Applications Key:  IF-F=Immunofluorescence (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-CREB (Ser133) (87G3) (Alexa Fluor® 488 Conjugate) Rabbit mAb detects endogenous levels of CREB only when phosphorylated at serine 133. The antibody also detects the phosphorylated form of the CREB-related protein, ATF-1.

Source / Purification

Monoclonal antibody is produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser133 of human CREB. The antibody was conjugated to Alexa Fluor® 488 under optimal conditions with an F/P ratio of 2-5.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of NIH/3T3 cells, untreated (blue) or PDGF treated (green), using Phospho-CREB (Ser133) (87G3) Rabbit mAb (Alexa Fluor® 488 Conjugate).

IF-IC

IF-IC

Confocal immunofluorescent analysis of SK-N-MC cells, treated with Forskolin and IBMX (left) or untreated (right), labeled with Phospho-CREB (Ser133) (87G3) Rabbit mAb (Alexa Fluor® 488 Conjugate) (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red).

IF-F

IF-F

Confocal immunofluorescent analysis of P14 rat cerebellum, untreated (left) or phosphatase treated (right), labeled with Phospho-CREB (Ser133) (87G3) Rabbit mAb (Alexa Fluor® 488 Conjugate) (green) and GFAP (GA5) Mouse mAb #3670 (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).


Description

Cell Signaling Technology Antibody conjugated to Alexa Fluor ® 488 fluorescent dye and tested in-house for direct Flow Cytometric analysis of human and mouse cells. The unconjugated antibody, #9198 reacts with Phospho-CREB (Ser133) from human, mouse and rat. CST expects that Phospho-CREB (Ser133) (87G3) Rabbit mAb (Alexa Fluor® 488 Conjugate) will also recognize Phospho-CREB in these species.

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, 371-385.
  2. Lee, M.M. et al. (1999) J. Neurosci. Res. 55, 702-712.
  3. Redmond, L. et al. (2002) Neuron 34, 999-1010.
  4. Dash, P.K. et al. (1990) Nature 345, 718-721.
  5. Yin, J.C. et al. (1994) Cell 79, 49-58.
  6. Guzowski, J.F. and McGaugh, J.L. (1997) Proc. Nat. Acad. Sci. USA 94, 2693-2698.
  7. Xing, J. et al. (1998) Mol. Cell. Biol. 18, 1946-1955.
  8. Ribar, T.J. et al. (2000) J. Neurosci. 20, RC107.
  9. Tan, Y. et al. (1996) EMBO J. 15, 4629-4642.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

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