Overview

  • Product nameAnti-Daxx antibody [E94]
    See all Daxx primary antibodies
  • Description
    Rabbit monoclonal [E94] to Daxx
  • Tested applicationsWB, IHC-P, ICC, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    A synthetic peptide corresponding to residues of human DAXX.

  • Positive control
    • WB: Hela cell lysate. IHC-P: Human stomach adenocarcinoma. IF/ICC: HeLa cell line.
  • General notes

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

Our Abpromise guarantee covers the use of ab32140 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 1/5000. Detects a band of approximately 100 kDa (predicted molecular weight: 81 kDa).
IHC-P Use at an assay dependent concentration.
ICC 1/50.
Flow Cyt 1/100.
  • Application notesIs unsuitable for IP.
  • Target

    • FunctionActs as an adapter protein in a MDM2-DAXX-USP7 complex by regulating the RING-finger E3 ligase MDM2 ubiquitination activity. Under non-stress condition, in association with the deubiquitinating USP7, prevents MDM2 self-ubiquitination and enhances the intrinsic E3 ligase activity of MDM2 towards TP53, thereby promoting TP53 ubiquitination and subsequent proteasomal degradation. Upon DNA damage, its association with MDM2 and USP7 is disrupted, resulting in increased MDM2 autoubiquitination and consequently, MDM2 degradation, which leads to TP53 stabilization. Proposed to mediate activation of the JNK pathway and apoptosis via MAP3K5 in response to signaling from TNFRSF6 and TGFBR2. Interaction with HSPB1/HSP27 may prevent interaction with TNFRSF6 and MAP3K5 and block DAXX-mediated apoptosis. In contrast, in lymphoid cells JNC activation and TNFRSF6-mediated apoptosis may not involve DAXX. Seems to regulate transcription in PML/POD/ND10 nuclear bodies together with PML and may influence TNFRSF6-dependent apoptosis thereby. Down-regulates basal and activated transcription. Seems to act as a transcriptional corepressor and inhibits PAX3 and ETS1 through direct protein-protein interaction. Modulates PAX5 activity. Its transcription repressor activity is modulated by recruiting it to subnuclear compartments like the nucleolus or PML/POD/ND10 nuclear bodies through interactions with MCSR1 and PML, respectively.
    • Tissue specificityUbiquitous.
    • Sequence similaritiesBelongs to the DAXX family.
    • Post-translational
      modifications
      Sumoylated.
      Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylated by HIPK1 upon glucose deprivation.
      Polyubiquitinated; which is promoted by CUL3 and SPOP and results in proteasomal degradation. Ubiquitinated by MDM2; inducing its degradation. Deubiquitinated by USP7; leading to stabilize it.
    • Cellular localizationCytoplasm. Nucleus > nucleoplasm. Nucleus > PML body. Nucleus > nucleolus. Chromosome > centromere. Dispersed throughout the nucleoplasm, in PML/POD/ND10 nuclear bodies, and in nucleoli. Colocalizes with a subset of interphase centromeres, but is absent from mitotic centromeres. Detected in cytoplasmic punctate structures. Translocates from the nucleus to the cytoplasm upon glucose deprivation or oxidative stress. Colocalizes with RASSF1 in the nucleus. Colocalizes with USP7 in nucleoplasma with accumulation in speckled structures.
    • Information by UniProt
    • Database links
    • Alternative names
      • antibody
      • BING 2 antibody
      • BING2 antibody
      • CENP-C binding protein antibody
      • DAP 6 antibody
      • DAP6 antibody
      • Daxx antibody
      • DAXX antibody
      • DAXX_HUMAN antibody
      • Death associated protein 6 antibody
      • Death domain associated protein 6 antibody
      • Death domain associated protein antibody
      • Death domain-associated protein 6 antibody
      • EAP 1 antibody
      • EAP1 antibody
      • ETS1 associated protein 1 antibody
      • ETS1-associated protein 1 antibody
      • Fas binding protein antibody
      • Fas death domain associated protein antibody
      • Fas death domain-associated protein antibody
      • hDaxx antibody
      • MGC126245 antibody
      • MGC126246 antibody
      see all

    Anti-Daxx antibody [E94] images

    • ab32140 stained HeLa cells. The cells were 4% formaldehyde fixed for 10 minutes at room temperature and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32140 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

    • Overlay histogram showing HeLa cells stained with ab32140 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32140, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • Anti-Daxx antibody [E94] (ab32140) at 1/5000 dilution + Hela cell lysate

      Predicted band size : 81 kDa
      Observed band size : 100 kDa (why is the actual band size different from the predicted?)
      Additional bands at : 48 kDa. We are unsure as to the identity of these extra bands.
    • Ab32140, at a dilution of 1/50, staining Daxx in paraffin embedded human stomach adenocarcinoma tissue by Immunohistochemistry.Ab32140, at a dilution of 1/50, staining Daxx in paraffin embedded human stomach adenocarcinoma tissue by Immunohistochemistry.

    References for Anti-Daxx antibody [E94] (ab32140)

    This product has been referenced in:
    • Full F  et al. Herpesvirus saimiri antagonizes nuclear domain 10-instituted intrinsic immunity via an ORF3-mediated selective degradation of cellular protein Sp100. J Virol 86:3541-53 (2012). Read more (PubMed: 22278248) »
    • Jiang M  et al. Functional reorganization of promyelocytic leukemia nuclear bodies during BK virus infection. MBio 2:e00281-10 (2011). WB . Read more (PubMed: 21304169) »

    See all 6 Publications for this product

    Product Wall

    Application Immunohistochemistry (Frozen sections)
    Blocking step 0.25%TritonX100, 0.2%Gelatine as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 25°C
    Sample Mouse Tissue sections (mouse embryonic spinal cord E11)
    Specification mouse embryonic spinal cord E11
    Permeabilization Yes - 0.25%TritonX100
    Fixative Paraformaldehyde
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    Submitted Aug 07 2014

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"