Revision 9

#7389

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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, W-S, IP, IHC-P, IF-IC, ChIP, ChIP-seq

Reactivity:
H M R Mk

Sensitivity:
Endogenous

MW (kDa):
300

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q92793

Entrez-Gene Id:
1387

Product Usage Information

For optimal ChIP 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.
Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 1:200
Immunohistochemistry (Paraffin) 1:50
Immunofluorescence (Immunocytochemistry) 1:100 - 1:200
Chromatin IP 1:50
Chromatin IP-seq 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #63794.

Specificity/Sensitivity

CBP (D6C5) Rabbit Monoclonal Antibody recognizes endogenous levels of total CBP protein. This antibody does not cross-react with p300 protein. Non-specific cytoplasmic staining was observed in cardiomyocytes by immunohistochemistry.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a recombinant protein specific to the amino terminus of human CBP protein. The epitope has been mapped to residues surrounding Ser235.

Background

CBP (CREB-binding protein) and p300 are highly conserved and functionally related transcriptional co-activators that associate with transcriptional regulators and signaling molecules, integrating multiple signal transduction pathways with the transcriptional machinery (1,2). CBP/p300 also contain histone acetyltransferase (HAT) activity, allowing them to acetylate histones and other proteins (2). Phosphorylation of p300 at Ser89 by PKC represses its transcriptional activity, and phosphorylation at the same site by AMPK disrupts the association of p300 with nuclear receptors (3,4). Ser1834 phosphorylation of p300 by Akt disrupts its association with C/EBPβ (5). Growth factors induce phosphorylation of CBP at Ser437, which is required for CBP recruitment to the transcription complex (6). CaM kinase IV phosphorylates CBP at Ser302, which is required for CBP-dependent transcriptional activation in the CNS (7). The role of acetylation of CBP/p300 is of particular interest (2,8). Acetylation of p300 at Lys1499 has been demonstrated to enhance its HAT activity and affect a wide variety of signaling events (9).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) ChIP: Chromatin IP ChIP-seq: Chromatin IP-seq

Cross-Reactivity Key

H: Human M: Mouse R: Rat Mk: Monkey

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 9

Cell Signaling Technology Logo
Western blot analysis of extracts from control HEK293T cells (Lane 1) or HEK293T cells with a targeted mutation in the gene encoding CBP (Lane 2) using CBP (D6C5) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The change in CBP molecular weight in the mutated HEK293T cells confirms the specificity of the antibody for CBP.
Western Blotting Image 1: CBP (D6C5) Rabbit Monoclonal Antibody
Western blot analysis of extracts from various cell lines using CBP (D6C5) Rabbit mAb.
Western Blotting Image 2: CBP (D6C5) Rabbit Monoclonal Antibody
Simple Western™ analysis of lysates (0.1 mg/mL) from 293T cells using CBP (D6C5) Rabbit mAb #7389. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 66 - 440 kDa separation module.
Western Blotting Image 1: CBP (D6C5) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 9

Cell Signaling Technology Logo
Immunoprecipitation of CBP from HeLa cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or CBP (D6C5) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using CBP (D6C5) Rabbit mAb.
Immunoprecipitation Image 1: CBP (D6C5) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using CBP (D6C5) Rabbit mAb.
Immunohistochemistry Image 1: CBP (D6C5) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human adenocarcinoma of the colon using CBP (D6C5) Rabbit mAb.
Immunohistochemistry Image 2: CBP (D6C5) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 9

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma using CBP (D6C5) Rabbit mAb.
Immunohistochemistry Image 3: CBP (D6C5) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded mouse ovary using CBP (D6C5) Rabbit mAb.
Immunohistochemistry Image 4: CBP (D6C5) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded normal human esophagus using CBP (D6C5) Rabbit mAb.
Immunohistochemistry Image 5: CBP (D6C5) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 9

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded LL2 syngeneic tumor using CBP (D6C5) Rabbit mAb.
Immunohistochemistry Image 6: CBP (D6C5) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded mouse colon using CBP (D6C5) Rabbit mAb.
Immunohistochemistry Image 7: CBP (D6C5) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded mouse cortex using CBP (D6C5) Rabbit mAb.
Immunohistochemistry Image 8: CBP (D6C5) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 9

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded mouse ovary using CBP (D6C5) Rabbit mAb.
Immunohistochemistry Image 9: CBP (D6C5) Rabbit Monoclonal Antibody
Immunohistochemical analysis of paraffin-embedded human adenoid cystic carcinoma of the salivary gland using CBP (D6C5) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 10: CBP (D6C5) Rabbit Monoclonal Antibody
Confocal immunofluorescent analysis of HeLa cells using CBP (D6C5) Rabbit mAb (green) and DyLight™ 554 Phalloidin #13054 (red).
Immunofluorescence Image 1: CBP (D6C5) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 9

Cell Signaling Technology Logo
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM, 1h) and CBP (D6C5) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across NR4A3, a known target gene of CBP (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
Chromatin Immunoprecipitation Image 1: CBP (D6C5) Rabbit Monoclonal Antibody
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells treated with Forskolin #3828 (30 μM, 1h) and CBP (D6C5) Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 9 (upper), including NR4A3 (lower), a known target gene of CBP (see additional figure containing ChIP-qPCR data).
Chromatin Immunoprecipitation Image 2: CBP (D6C5) Rabbit Monoclonal Antibody
Chromatin immunoprecipitations were performed with cross-linked chromatin from 293 cells, treated with Forskolin #3828 (30 μM, 1h) and either CBP (D6C5) Rabbit mAb or 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.
Chromatin Immunoprecipitation Image 3: CBP (D6C5) Rabbit Monoclonal Antibody
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.