Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-CREB (Ser133) (87G3) Rabbit mAb #9198

Applications Reactivity Sensitivity MW (kDa) Isotype
W IHC-P IHC-F IF-F IF-IC F ChIP H M R (Z) Endogenous 43 Rabbit IgG

Applications Key:  W=Western Blotting  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-F=Immunofluorescence (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Z=Zebrafish
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-CREB (Ser133) (87G3) 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 animals with a synthetic phosphopeptide corresponding to residues surrounding Ser133 of human CREB.

Western Blotting

Western Blotting

Western blot analysis of extracts from SK-N-MC cells, untreated or forskolin- and FGF-treated, using Phospho-CREB (Ser133) (87G3) Rabbit mAb (upper) or CREB (48H2) Rabbit mAb #9197 (lower).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, showing nuclear staining, using Phospho-CREB (Ser133) (87G3) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded mouse lung using Phospho-CREB (Ser133) (87G3) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using Phospho-CREB (Ser133) (87G3) Rabbit mAb in the presence of control peptide (left) or Phospho-CREB (Ser133) Blocking Peptide #1090 (right).

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded SK-N-MC cells, untreated (left) or IBMX- and forskolin-treated (right), showing induced nuclear staining, using Phospho-CREB (Ser133) (87G3) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma, untreated (left) or lambda phosphatase-treated (right), using Phospho-CREB (Ser133) (87G3) Rabbit mAb.


IHC-F (frozen)

IHC-F (frozen)

Immunohistochemical analysis of frozen H1650 xenograft, showing nuclear localization using Phospho-CREB (Ser133)(87G3) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of SK-N-MC cells, untreated (blue) or IBMX- and forskolin-treated (green), using Phospho-CREB (Ser133) (87G3) Rabbit mAb compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal microscopic images of SK-N-MC cells showing nuclear stain after 25 minute treatment with Forskolin and IBMX using Phospho-CREB (Ser133) (87G3) Rabbit mAb (left, red) compared to untreated cells (right). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


IF-F

IF-F

Conofocal immunofluorescent images of rat dentate gyrus, either sham-operated (left) or 15 min ischemia followed by 30 min (center) and 4 h (right) reperfusion, labeled with Phospho-CREB (Ser133) (87G3) Rabbit mAb (red), Neurofilament-L (DA2) Mouse mAb #2835 (blue) and Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb (Alexa Fluor® 488 Conjugate) #4854.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293 cells treated with Forskolin #3828 (30 μM) for 1h and either 20 μl of Phospho-CREB (Ser133) (87G3) Rabbit mAb 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 human ALS2 exon 1 primers, SimpleChIP® Human NR4A3 Promoter Primers #4829, and SimpleChIP® Human α Satellite Repeat Primers #4486. 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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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

For Research Use Only. Not For Use In Diagnostic Procedures.

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