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RecombinantRabMAb

Recombinant Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)

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Western blot - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Immunocytochemistry/ Immunofluorescence - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Flow Cytometry - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Immunoprecipitation - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Western blot - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Western blot - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] (ab253202)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Immunocytochemistry/ Immunofluorescence - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Flow Cytometry - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • Immunoprecipitation - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • ChIP - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
  • 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

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • 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. 

  • 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.

  • 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.

  • 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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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
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.
Notes
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

  • 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
    modifications

    Methylation 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

  • Database links

    see all

  • Alternative names

    • CBP antibody
    • CBP_HUMAN antibody
    • CREB binding protein antibody
    • CREB-binding protein antibody
    • Crebbp antibody
    • Cyclic AMP responsive enhancer binding protein antibody
    • KAT3A antibody
    • RSTS antibody
    • RTS antibody
    • Rubinstein Taybi syndrome antibody
    see all

Images

  • Western blot - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    Western blot - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    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.

  • Flow Cytometry - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    Flow Cytometry - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)

    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.

  • Immunoprecipitation - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    Immunoprecipitation - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)

    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.

  • Western blot - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    Western blot - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    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.

     

  • Western blot - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    Western blot - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    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.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] (ab253202)
    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    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)
    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)
    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.

  • Flow Cytometry - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    Flow Cytometry - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)

    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.

  • Immunoprecipitation - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    Immunoprecipitation - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)

    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.

  • ChIP - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    ChIP - Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)

    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

  • Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)
    Anti-CREBBP antibody [EPR23418-23] - ChIP Grade (ab253202)

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.

Click here to view the general protocols

References (0)

Publishing research using ab253202? Please let us know so that we can cite the reference in this datasheet.

ab253202 has not yet been referenced specifically in any publications.

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