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Western Blot analysis of extracts from SK-N-MC, COS, NIH/3T3, C6 and Drosophila S2 cells, using CREB (48H2) Rabbit mAb.

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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).

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After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

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Immunohistochemical analysis of paraffin-embedded human astrocytoma, using CREB (48H2) Rabbit mAb.

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

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Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using CREB (48H2) Rabbit mAb.

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

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Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma, using CREB (48H2) Rabbit mAb.

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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).

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Immunohistochemical analysis of paraffin-embedded mouse brain, using CREB (48H2) Rabbit mAb.

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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.

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Immunohistochemical analysis of frozen H1650 xenograft, showing nuclear localization using CREB (48H2) Rabbit mAb.

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

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Flow cytometric analysis of SK-N-MC cells using CREB (48H2) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

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

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Confocal immunofluorescent analysis of SK-N-MC cells showing nuclear stain with CREB (48H2) Rabbit mAb (A, red) compared to an isotype control (B). Blue pseudocolor =DRAQ5® (fluorescent DNA dye).

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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).

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Confocal immunofluorescent analysis of mouse cerebellum labeled with CREB (48H2) Rabbit mAb (red) and Neurofilament-L (DA2) Mouse mAb #2835 (green). Blue pseudocolor =DRAQ5® #4084 (fluorescent DNA dye).

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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).

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Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293 cells, treated with Forskolin #3828 (30 μM) for 1h and either 10 μl of CREB (48H2) 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.

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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.

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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.

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Image
Product Includes Quantity Applications Reactivity MW(kDa) Isotype
CREB (48H2) Rabbit mAb 9197 100 µl
H M R Mk Dm 43 Rabbit IgG
Phospho-CREB (Ser133) (87G3) Rabbit mAb 9198 100 µl
H M R 43 Rabbit IgG
CREB Control Cell Extracts 9193 100 µl
 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
All Goat 
Anti-biotin, HRP-linked Antibody 7075 100 µl
Goat 
20X LumiGLO® Reagent and 20X Peroxide 7003 5 ml each
 
Biotinylated Protein Ladder Detection Pack 7727 100 µl
 

Product Description

The PhosphoPlus CREB (Ser133) Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of CREB at Ser133.


Specificity / Sensitivity

Phospho-CREB (Ser133) (87G3) Rabbit mAb detects endogenous levels of CREB only when phosphorylated at Ser133. This antibody also detects the phosphorylated form of CREB-related protein ATF-1. CREB (48H2) Rabbit mAb detects endogenous levels of total CREB protein.


Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser133 of human CREB (Phospho-CREB (Ser133) (87G3) Rabbit mAb), or with a synthetic peptide corresponding to residues near the carboxy-terminus of human CREB (CREB (48H2) Rabbit mAb).

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.


Entrez-Gene Id 1385
Swiss-Prot Acc. P16220


For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
PhosphoPlus® is a trademark of Cell Signaling Technology, Inc.
DRAQ5® is a registered trademark of Biostatus Limited.
LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.
U.S. Patent No. 5,675,063.