Overview

  • Product name
    Anti-TRAF6 antibody [EP591Y] - BSA and Azide free
    See all TRAF6 primary antibodies
  • Description
    Rabbit monoclonal [EP591Y] to TRAF6 - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Zebrafish
  • Immunogen

    A synthetic peptide corresponding to residues near the N term of TRAF6 (Human)

  • Positive control
    • Jurkat cell lysate, Human colon adenocarcinoma
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    ab218575 is a PBS-only buffer formulated version of ab33915, containing no BSA or sodium azide, ideal for antibody labeling. Please refer to ab33915 for information on protocols, dilutions, and image data.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab218575 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
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB Use at an assay dependent concentration. Predicted molecular weight: 58 kDa.Can be blocked with TRAF6 peptide (ab183540).
IHC-P Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      E3 ubiquitin ligase that, together with UBE2N and UBE2V1, mediates the synthesis of 'Lys-63'-linked-polyubiquitin chains conjugated to proteins, such as IKBKG, AKT1 and AKT2. Also mediates ubiquitination of free/unanchored polyubiquitin chain that leads to MAP3K7 activation. Leads to the activation of NF-kappa-B and JUN. May be essential for the formation of functional osteoclasts. Seems to also play a role in dendritic cells (DCs) maturation and/or activation. Represses c-Myb-mediated transactivation, in B lymphocytes. Adapter protein that seems to play a role in signal transduction initiated via TNF receptor, IL-1 receptor and IL-17 receptor.
    • Tissue specificity
      Expressed in heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.
    • Pathway
      Protein modification; protein ubiquitination.
    • Sequence similarities
      Belongs to the TNF receptor-associated factor family. A subfamily.
      Contains 1 MATH domain.
      Contains 1 RING-type zinc finger.
      Contains 2 TRAF-type zinc fingers.
    • Domain
      The coiled coil domain mediates homo- and hetero-oligomerization.
      The MATH/TRAF domain binds to receptor cytoplasmic domains.
    • Post-translational
      modifications
      Sumoylated on Lys-124, Lys-142 and Lys-453 by SUMO1.
      Polyubiquitinated on Lys-124; after cell stimulation with IL-1-beta or TGF-beta. This ligand-induced cell stimulation leads to dimerization/oligomerization of TRAF6 molecules, followed by auto-ubiquitination which involves UBE2N and UBE2V1 and leads to TRAF6 activation. This 'Lys-63' site-specific poly-ubiquitination appears to be associated with the activation of signaling molecules. Endogenous autoubiquitination occurs only for the cytoplasmic form.
    • Cellular localization
      Cytoplasm. Cytoplasm > cell cortex. Nucleus. Found in the nuclei of some agressive B-cell lymphoma cell lines as well as in the nuclei of both resting and activated T-and B-lymphocytes. Found in punctate nuclear body protein complexes. Ubiquitination may occur in the cytoplasm and sumoylation in the nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • E3 ubiquitin-protein ligase TRAF6 antibody
      • Interleukin 1 signal transducer antibody
      • Interleukin-1 signal transducer antibody
      • MGC 3310 antibody
      • MGC:3310 antibody
      • MGC3310 antibody
      • OTTHUMP00000232772 antibody
      • OTTHUMP00000232773 antibody
      • RING finger protein 85 antibody
      • RNF 85 antibody
      • RNF85 antibody
      • TNF receptor associated factor 6 antibody
      • TNF receptor-associated factor 6 antibody
      • TNF receptor-associated factor 6, E3 ubiquitin protein ligase antibody
      • TRAF 6 antibody
      • Traf6 antibody
      • TRAF6_HUMAN antibody
      see all

    Images

    • This WB data was generated using the same anti-TRAF6 antibody clone, EP591Y, in a different buffer formulation (cat# ab33915).

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)              
      Lane 2: TRAF6 knockout  HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: HEK293 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab33915 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.

      Ab33915 was shown to specifically react with TRAF6 in wild-type cells as signal was lost in TRAF6 knockout HAP1 cells. Wild-type and TRAF6 knockout samples were subjected to SDS-PAGE.  Ab33915 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 2000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging

    • Overlay histogram showing HAP1 wildtype (green line) and HAP1-TRAF6 knockout cells (red line) stained with ab33915. The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab33915, 0.1µg/ml ) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) preadsorbed (ab150081) at 1/2000 dilution for 30 min at 22°C.

      A rabbit IgG isotype control antibody  (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-TRAF6  knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

      Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33915).

    • Flow Cytometry analysis of Jurkat (human acute T cell leukemia) cells labeling TRAF6 with purified ab33915 at 1/240 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33915).

    • ab33915 staining TRAF6 in Human platelet cells by Flow cytometry.
      Cells were fixed in paraformaldehyde and permeabilized using 0.1% Triton-X-100 in 2% BSA for 15 minutes. Primary antibody used at a 1/200 dilution and incubated for 16 hours at 4°C. The secondary antibody used was an Alexa Fluor®488 conjugated chicken anti-rabbit IgG (H+L) at a 1/500 dilution.

      P : Permeabilized
      US : Unstained (Red Peak)
      IGG RB : IgG Rabbit (Blue Peak)
      TRAF6 Ab (Green Peak)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab33915).

    • This IHC data was generated using the same anti-TRAF6 antibody clone, EP591Y, in a different buffer formulation (cat# ab33915).

      Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using anti-TRAF6 (ab33915)

    References

    This product has been referenced in:
    • Meininger I  et al. Alternative splicing of MALT1 controls signalling and activation of CD4(+) T cells. Nat Commun 7:11292 (2016). Mouse . Read more (PubMed: 27068814) »
    • Padrão AI  et al. Endurance training prevents TWEAK but not myostatin-mediated cardiac remodelling in cancer cachexia. Arch Biochem Biophys 567:13-21 (2015). Read more (PubMed: 25575785) »
    See all 12 Publications for this product

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

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