CREB (48H2) Rabbit mAbProduct information
Product Pathways - Neuroscience
CREB (48H2) Rabbit mAb #9197
|9197L||300 µl (30 western blots)||---||In Stock||---|
|9197S||100 µl (10 western blots)||---||In Stock||---|
|9197T||20 µl (2 western blots)||---||In Stock||---|
|9197||carrier free and custom formulation / quantity||email request|
|W||1:1000||Human, Mouse, Rat, Monkey, D. melanogaster||Endogenous||43||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), IHC-F=Immunohistochemistry (Frozen), IF-F=Immunofluorescence (Frozen), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP, ChIP-seq=Chromatin IP-seq
Directions For Use
For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
Specificity / Sensitivity
CREB (48H2) Rabbit mAb detects endogenous levels of total CREB-1 protein. The antibody does not cross-react with other ATF/CREB family members.
Source / Purification
Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human CREB-1 protein.
Western Blot analysis of extracts from SK-N-MC, COS, NIH/3T3, C6 and Drosophila S2 cells, using CREB (48H2) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human astrocytoma, using CREB (48H2) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using CREB (48H2) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma, using CREB (48H2) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse brain, using CREB (48H2) Rabbit mAb.
Immunohistochemical analysis of frozen H1650 xenograft, showing nuclear localization using CREB (48H2) Rabbit mAb.
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).
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).
Flow cytometric analysis of SK-N-MC cells using CREB (48H2) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293 cells treated with Forskolin #3828 (30 μM) for 1h and 10 μl of CREB (48H2) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. The figure shows binding across NR4A3, a known target gene of CREB (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
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.
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).
- Lonze, B.E. et al. (2002) Neuron 34, 371-85.
- Lee, M.M. et al. (1999) J Neurosci Res 55, 702-12.
- Redmond, L. et al. (2002) Neuron 34, 999-1010.
- Dash, P.K. et al. (1990) Nature 345, 718-21.
- Yin, J.C. et al. (1994) Cell 79, 49-58.
- Guzowski, J.F. and McGaugh, J.L. (1997) Proc Natl Acad Sci USA 94, 2693-8.
- Xing, J. et al. (1998) Mol Cell Biol 18, 1946-55.
- Ribar, T.J. et al. (2000) J Neurosci 20, RC107.
- Tan, Y. et al. (1996) EMBO J 15, 4629-42.
- Banno, Y. et al. (2008) J Neurochem 104, 1372-86. Applications: Western Blotting.
- Kumar, A.P. et al. (2007) Clin Cancer Res 13, 2784-94. Applications: IHC-P (paraffin).
- Ghosh, R. et al. (2007) Neoplasia 9, 893-9. Applications: IHC-P (paraffin).
- Sakaguchi, M. et al. (2008) Mol Biol Cell 19, 78-85. Applications: Western Blotting.
- Gaddini, L. et al. (2009) Neurobiol Dis 35, 278-85. Applications: Western Blotting.
- Almeida, L.E. et al. (2009) J Neurosci 29, 12702-10. Applications: Western Blotting.
- Gehani, S.S. et al. (2010) Mol Cell 39, 886-900. Applications: Western Blotting.
- Chung, Y.W. et al. (2011) J Biol Chem 286, 29681-90. Applications: Western Blotting.
- Mihaylova, M.M. et al. (2011) Cell 145, 607-21. Applications: Western Blotting.
- Kir, S. et al. (2014) Nature , . Applications: Western Blotting.
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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.
Ultra is a trademark of New England Biolabs, Inc.
Illumina is a registered trademark of Illumina, Inc.
DRAQ5 is a registered trademark of Biostatus Limited.
NEBNext is a registered trademark of New England Biolabs, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.