CREB (48H2) Rabbit mAb
Product information| Code | Name | Size | Quantity | Price | |
|---|---|---|---|---|---|
9197T |
CREB (48H2) Rabbit mAb |
20 µl (2 western blots) | - | Unavailable in your region | |
9197S |
CREB (48H2) Rabbit mAb |
100 µl (10 western blots) | - | Unavailable in your region | |
9197L |
CREB (48H2) Rabbit mAb |
300 µl (30 western blots) | - | Unavailable in your region |
Product Pathways - Neuroscience
CREB (48H2) Rabbit mAb #9197
PhosphoSitePlus® protein, site, and accession data:
CREB
| No. | Size | Price | ||
|---|---|---|---|---|
| 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 | ||
| Application | Dilution | Species-Reactivity | Sensitivity | MW (kDa) | Isotype |
|---|---|---|---|---|---|
| W | 1:1000 | Human, Mouse, Rat, Monkey, D. melanogaster | Endogenous | 43 | Rabbit IgG |
| IP | 1:250 | ||||
| IHC-P | 1:6400 | ||||
| IHC-F | 1:6400 | ||||
| IF-F | 1:800 | ||||
| IF-IC | 1:800 | ||||
| F | 1:400 | ||||
| ChIP | 1:50 | ||||
| ChIP-seq | 1:50 |
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.
Protocols
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 Blotting
Western Blot analysis of extracts from SK-N-MC, COS, NIH/3T3, C6 and Drosophila S2 cells, using CREB (48H2) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human astrocytoma, using CREB (48H2) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using CREB (48H2) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin's lymphoma, using CREB (48H2) Rabbit mAb.
IHC-P (paraffin)
Immunohistochemical analysis of paraffin-embedded mouse brain, using CREB (48H2) Rabbit mAb.
IHC-F (frozen)
Immunohistochemical analysis of frozen H1650 xenograft, showing nuclear localization using CREB (48H2) Rabbit mAb.
IF-F
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).
IF-IC
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 Cytometry
Flow cytometric analysis of SK-N-MC cells using CREB (48H2) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
Chromatin IP-seq
Chromatin immunoprecipitations were performed with cross-linked chromatin from 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 IP
Chromatin immunoprecipitations were performed with cross-linked chromatin from 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.
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).
- 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.
Application References
- 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.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
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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.
The supporting documents available for this product can be downloaded below.