Product Pathways - Neuroscience

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

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Applications	Reactivity	Sensitivity	MW (kDa)	Isotype
W IHC-P IHC-F IF-F IF-IC F ChIP	H M R	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
Species cross-reactivity is determined by Western blot.

Protocols

9198:: ChIP Agarose, ChIP Magnetic, Flow, IHC / Frozen, IHC / Paraffin, Immunofluorescence, Western Blotting

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 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)

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

IHC-P (paraffin)

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

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)

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)

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)

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

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

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

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 immunoprecipitations were performed with cross-linked chromatin from 4 x 10⁶ 293 cells treated with Forskolin #3828 (30uM) for 1h and either 20 μl of Phospho-CREB (Ser133) (87G3) Rabbit mAb #9198 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).

Application References

Zaru, R. et al. (2007) Nat Immunol 8, 1227-35. Applications: Western Blotting
Ghosh, R. et al. (2007) Neoplasia 9, 893-9. Applications: IHC-P (paraffin)
Kumar, A.P. et al. (2007) Clin Cancer Res 13, 2784-94. Applications: IHC-P (paraffin)
Gaddini, L. et al. (2009) Neurobiol Dis 35, 278-85. Applications: IF-IC (In Cells) Western Blotting

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Companion Products

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

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.