Overview

  • Product name

    Anti-CREBBP antibody [CBP5I001]
    See all CREBBP primary antibodies
  • Description

    Mouse monoclonal [CBP5I001] to CREBBP
  • Host species

    Mouse
  • Tested applications

    Suitable for: Dot blot, IHC-P, WB, IP, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant full length protein corresponding to Human CREBBP.

  • Positive control

    • HeLa whole cell lysate (ab150035); NIH3T3 whole cell lysate. IHC-P: Human pancreas FFPE tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab50702 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
Dot blot Use at an assay dependent concentration.
IHC-P Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/50 - 1/100. Predicted molecular weight: 265 kDa.
IP 1/30.
ICC/IF 1/5.

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

  • Lane 1: Wild-type HAP1 whole cell lysate (40 µg)
    Lane 2: CREBBP knockout HAP1 whole cell lysate (40 µg)
    Lane 3: HeLa whole cell lysate (40 µg)
    Lane 4: NIH3T3 whole cell lysate (40 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab50702 observed at 265 kDa. Red - loading control, ab176560, observed at 50 kDa. 

    ab50702 was shown to specifically recognize CREBBP in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when CREBBP knockout samples were examined. Wild-type and CREBBP knockout samples were subjected to SDS-PAGE. Ab50702 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 1/100 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.              

  • IHC image of CREBBP staining in human pancreas formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab50702, 10µg/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.

  • Staining of HeLa cell lysate with ab50702 at a dilution of 1/5 and Goat anti-mouse secondary at 1/200.
  • Anti-CREBBP antibody [CBP5I001] (ab50702) at 1/30 dilution + RIPA lysate of HeLa cells at 500 µg

    Secondary
    Mouse IgG antibody at 1/2500 dilution

    Predicted band size: 265 kDa

  • Anti-CREBBP antibody [CBP5I001] (ab50702) at 1/50 dilution + NIH3T3 whole cell lysate at 50 µg

    Secondary
    Mouse IgG antibody at 1/2500 dilution

    Predicted band size: 265 kDa

  • Anti-CREBBP antibody [CBP5I001] (ab50702) at 1/100 dilution + HeLa whole cell lysate at 50 µg

    Secondary
    Mouse IgG antibody at 1/2500 dilution

    Predicted band size: 265 kDa

References

This product has been referenced in:

  • Li P  et al. Farnesoid X receptor interacts with cAMP response element binding protein to modulate glucagon-like peptide-1 (7-36) amide secretion by intestinal L cell. J Cell Physiol 234:12839-12846 (2019). Read more (PubMed: 30536761) »
  • Dong H  et al. SP1-induced lncRNA AGAP2-AS1 expression promotes chemoresistance of breast cancer by epigenetic regulation of MyD88. J Exp Clin Cancer Res 37:202 (2018). Read more (PubMed: 30157918) »
See all 4 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you for submitting an Abreview for ab50702. As you may have noticed, your review has now been published on our website.

Since you obtained poor results using the antibody in an untested application, we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.

Based on the information in your review, I feel that altering the following step in your protocol may help improve your results:
- incubate longer with the primary Ab (e.g. overnight at 4 degree)
- use other antigen retrieval than the one you tried (e.g. enzymatic)


If you decide to follow these suggestions, please let me know if they are helpful.
I wish you good luck with your research.

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (bladder)
Specification
bladder
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C

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Verified customer

Submitted Oct 30 2012

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