Anti-KAT3A / CBP antibody - ChIP Grade (ab2832)

Overview

  • Product name
    Anti-KAT3A / CBP antibody - ChIP Grade
    See all KAT3A / CBP primary antibodies
  • Description
    Rabbit polyclonal to KAT3A / CBP - ChIP Grade
  • Tested applications
    Suitable for: WB, ChIP, IHC-P, IP, ICC/IF, CHIPseq, Inhibition Assaymore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Non human primates
  • Immunogen

    Synthetic peptide corresponding to Human KAT3A/ CBP aa 162-176.
    Sequence:

    ATSSPATSQTGPGIC


    (Peptide available as ab4916)

  • Positive control
    • HeLa cell lysate. IHC-P: human pancreas FFPE tissue sections

Properties

Applications

Our Abpromise guarantee covers the use of ab2832 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
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 265 kDa (predicted molecular weight: 265 kDa).Can be blocked with Human KAT3A / CBP peptide (ab4916).
ChIP Use at an assay dependent concentration. PubMed: 20111703

Recommended to be used at 2-4µg for 1-2x106 cells.

IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP Use at an assay dependent concentration. PubMed: 20403362
ICC/IF 1/100.
CHIPseq Use at an assay dependent concentration. PubMed: 22196736
Inhibition Assay Use at an assay dependent concentration.

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
  • 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

  • Sonicated chromatin prepared from U2OS cells was subjected to the ChIP procedure with ab2832 to CBP. Immunoprecipitated chromatin was analysed in the promoter region of c-FOS (active) and in exon 2 of MYO-D (inactive). Values are % of inputs.  2–4 µg of ab2832 and 1-2x106 cells were used in each ChIP experiment.

    Sergei Denissov, Prof. Henk Stunnenberg lab, University Nijmengen, NL

  • Immunofluorescent analysis of MCF-7 cells, labeling KAT3A/CBP with ab2832 (right) compared with a negative control without ab2832 (left). Cells were formalin fixed, permeabilized with 0.1% Triton X-100 for 5-10 minutes, and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with anti-KAT3A/CBP diluted 1/100 in 3% BSA/PBS overnight at 4°C. Actin was stained using Alexa Fluor 554 (red) and nuclei were stained with DAPI (blue). The nuclear localization of KAT3A/CBP can be observed (green).

  • IHC image of KAT3A/CBP staining in human pancreas formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2832, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • Immunofluorescent analysis of HeLa cells, labeling KAT3A/CBP with ab2832 (right) compared with a negative control without ab2832 (left). Cells were formalin fixed, permeabilized with 0.1% Triton X-100 for 5-10 minutes, and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with anti-KAT3A/CBP diluted 1/100 in 3% BSA/PBS overnight at 4°C. Actin was stained using Alexa Fluor 554 (red) and nuclei were stained with DAPI (blue). The nuclear localization of KAT3A/CBP can be observed (green). 

  • Immunohistochemical analysis of FFPE mouse colon tissue, labeling KAT3A/CBP with ab2832 (right) compared with a negative control without ab2832 (left). Heat antigen retrieval performed with 10mM Sodium Citrate (pH 6) for 8-15 minutes. Tissue blocked with 3% H2O2-methanol for 15 minutes at room temperature. Incubation with ab2832 diluted 1/2000 in 3% BSA-PBS overnight at 4°C. Counterstaining with hematoxylin. 

  • Immunofluorescent analysis of NIH-3T3 cells, labeling KAT3A/CBP with ab2832 (right) compared with a negative control without ab2832 (left). Cells were formalin fixed, permeabilized with 0.1% Triton X-100 for 5-10 minutes, and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were incubated with anti-KAT3A/CBP diluted 1/100 in 3% BSA/PBS overnight at 4°C. Nuclei were stained with DAPI (blue).



  • Predicted band size : 265 kDa
    ab2832 using HeLa cell lysate. ab2832 using HeLa cell lysate.

References

This product has been referenced in:
  • Tsukamoto D  et al. HNF-4 participates in the hibernation-associated transcriptional regulation of the chipmunk hibernation-related protein gene. Sci Rep 7:44279 (2017). ChIP . Read more (PubMed: 28281641) »
  • Liu Y  et al. Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis. Respir Res 18:38 (2017). WB ; Rat . Read more (PubMed: 28222740) »

See all 29 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole brain tissue lysate)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (3-8% Tris-Acetate)
Loading amount
20 µg
Specification
Whole brain tissue lysate
Blocking step
BSA as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Mar 31 2017

Application
ChIP
Detection step
Real-time PCR
Sample
Mouse Tissue lysate - nuclear (Liver)
Specification
Liver
Negative control
no ab
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: PFA 1%
Positive control
ChIP with anti-Pol ab
Username

Abcam user community

Verified customer

Submitted Sep 16 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Heat-inactivated normal donkey serum in 0.05% PBS-T as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Antigen retrieval step
None
Sample
Mouse Tissue sections (Mouse, whole brain sections)
Specification
Mouse, whole brain sections
Permeabilization
Yes - Tween-20
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 13 2014

Application
Western blot
Sample
Chicken Tissue lysate - whole (brain)
Loading amount
5.2 µg
Specification
brain
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Ms. Ying Jiang

Verified customer

Submitted Sep 21 2012

Thank you for your enquiry.
The exact immunogen sequence used to generate ab2832 is noted on the online product datasheet as:
Synthetic peptide: ATSSPATSQTGPGIC, corresponding to amino acids 162-176 in the nuclear factor binding domain of Hum...

Read More

Thank you for contacting us. We have the following antibodies in our catalog that where tested in ChIP: for CBP: ab2832, ab10489, and ab10490 for p300: ab14984 None of the antibodies has been tested so far in zebrafish. If you ...

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