Recombinant Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23418-23] to CREBBP - ChIP Grade
- Suitable for: IHC-P, WB, ICC/IF, ChIP, Flow Cyt (Intra), IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-CREBBP antibody [EPR23418-23] - ChIP Grade
See all CREBBP primary antibodies -
Description
Rabbit monoclonal [EPR23418-23] to CREBBP - ChIP Grade -
Host species
Rabbit -
Specificity
This antibody does not react with mouse species for ChIP application.
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Tested applications
Suitable for: IHC-P, WB, ICC/IF, ChIP, Flow Cyt (Intra), IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HeLa, HeLa, 293T, NIH/3T3 and PC-12 whole cell lysate; His-tagged human CREBBP recombinant protein. IHC-P: Human bladder cancer, Human cervical cancer, Mouse cerebrum and Rat cerebrum tissue. ICC: HeLa and NIH/3T3 cells. Flow Cyt: HeLa and NIH/3T3 cells. IP: NIH/3T3 and HeLa cells. ChIP: HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR23418-23 -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab253202 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB |
1/1000. Predicted molecular weight: 51 kDa.
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ICC/IF |
1/100.
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ChIP |
Use a concentration of 5 µg/ml.
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Flow Cyt (Intra) |
1/500.
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IP |
1/30.
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Notes |
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IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 51 kDa. |
ICC/IF
1/100. |
ChIP
Use a concentration of 5 µg/ml. |
Flow Cyt (Intra)
1/500. |
IP
1/30. |
Target
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Function
Acetylates histones, giving a specific tag for transcriptional activation. Also acetylates non-histone proteins, like NCOA3 coactivator. Binds specifically to phosphorylated CREB and enhances its transcriptional activity toward cAMP-responsive genes. Acts as a coactivator of ALX1 in the presence of EP300. -
Involvement in disease
Note=Chromosomal aberrations involving CREBBP may be a cause of acute myeloid leukemias. Translocation t(8;16)(p11;p13) with MYST3/MOZ; translocation t(11;16)(q23;p13.3) with MLL/HRX; translocation t(10;16)(q22;p13) with MYST4/MORF. MYST3-CREBBP may induce leukemia by inhibiting RUNX1-mediated transcription.
Defects in CREBBP are a cause of Rubinstein-Taybi syndrome type 1 (RSTS1) [MIM:180849]. RSTS1 is an autosomal dominant disorder characterized by craniofacial abnormalities, broad thumbs, broad big toes, mental retardation and a propensity for development of malignancies. -
Sequence similarities
Contains 1 bromo domain.
Contains 1 KIX domain.
Contains 2 TAZ-type zinc fingers.
Contains 1 ZZ-type zinc finger. -
Domain
The KIX domain mediates binding to HIV-1 Tat. -
Post-translational
modificationsMethylation of the KIX domain by CARM1 blocks association with CREB. This results in the blockade of CREB signaling, and in activation of apoptotic response.
Phosphorylated upon DNA damage, probably by ATM or ATR.
Sumoylation negatively regulates transcriptional activity via the recruitment of DAAX. -
Cellular localization
Cytoplasm. Nucleus. Recruited to nuclear bodies by SS18L1/CREST. In the presence of ALX1 relocalizes from the cytoplasm to the nucleus. - Information by UniProt
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Database links
- Entrez Gene: 1387 Human
- Entrez Gene: 12914 Mouse
- Entrez Gene: 54244 Rat
- Omim: 600140 Human
- SwissProt: Q92793 Human
- SwissProt: P45481 Mouse
- SwissProt: Q6JHU9 Rat
- Unigene: 459759 Human
see all -
Alternative names
- CBP antibody
- CBP_HUMAN antibody
- CREB binding protein antibody
see all
Images
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All lanes : Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202) at 1/1000 dilution
Lane 1 : HAP1 (human chronic myelogenous leukemia near-haploid cell), whole cell lysate at 28 µg
Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 14 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 51 kDa
Observed band size: 300 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST. Fresh lysates were used in this WB.
Exposure time: Lane 1: 81 seconds; Lane 2: 3 minutes.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
Immunohistochemical analysis of paraffin-embedded Human bladder cancer tissue labelling CREBBP with ab253202 at 1/2000 (0.258 µg/mL) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining in human bladder cancer (PMID: 25915404). The section was incubated with ab253202 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunocytochemistry/ Immunofluorescence - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa cells labelling CREBBP with ab253202 at 1/500 (1.032 µg/mL) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/mL) dilution (Green). Confocal image showing mianly nuclear staining in HeLa cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 µg/mL) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling CREBBP with ab253202 at 1/500 dilution (0.1µg)/ red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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CREBBP was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 µg with ab253202 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab253202 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate 10 ug
Lane 2: ab253202 IP in NIH/3T3 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab253202 in NIH/3T3 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 110 seconds.
Fresh lysates were used in this IP.
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All lanes : Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : 293T (human embryonic kidney epithelial cell), whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 51 kDa
Observed band size: 300 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST. Fresh lysates were used in this WB.
Exposure time: 81 seconds.
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All lanes : Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202) at 1/1000 dilution
Lane 1 : His-tagged human CREBBP recombinant protein (aa2221-2442)
Lane 2 : His-tagged human EP300 recombinant protein (aa2215-2414)
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 51 kDa
Observed band size: 25 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST. This antibody has no cross-reaction with human EP300.
These two recombinant proteins were made in-house and expressed from the E.coli expression system.
Exposure time: 1 second.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] (ab253202)
Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labelling CREBBP with ab253202 at 1/2000 (0.258 µg/mL) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining in human cervical cancer. The section was incubated with ab253202 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling CREBBP with ab253202 at 1/2000 (0.258 µg/mL) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Mainly nuclear staining in mouse cerebrum. The section was incubated with ab253202 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling CREBBP with ab253202 at 1/2000 (0.258 µg/mL) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Mainly nuclear staining in rat cerebrum (PMID: 12445700). The section was incubated with ab253202 for 10 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
-
Immunocytochemistry/ Immunofluorescence - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labelling CREBBP with ab253202 at 1/100 (5.16 µg/mL) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 µg/mL) dilution (Green). Confocal image showing mianly nuclear staining in NIH/3T3 cell line is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 µg/mL) dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/mL) dilution.
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Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling CREBBP with ab253202 at 1/500 dilution (0.1µg)/ red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
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CREBBP was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate 10 µg with ab253202 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab253202 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP) (ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg
Lane 2: ab253202 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab253202 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 48 seconds.
Fresh lysates were used in this IP.
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Chromatin was prepared from HeLa cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab253202 (red), or 5 µg of rabbit normal IgG ab172730 (gray) and 25 µl of Protein A/G Dynabeads. The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci).
Primers are from paper PMID:2178921
* http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab253202 has not yet been referenced specifically in any publications.