Overview

  • Product name
    Anti-TRAF6 antibody [EP592Y] - BSA and Azide free
    See all TRAF6 primary antibodies
  • Description
    Rabbit monoclonal [EP592Y] to TRAF6 - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human TRAF6 aa 500 to the C-terminus (C terminal).

  • Positive control
    • WB: Jurkat, HEK293 and HeLa cell lysates. IHC-P: Human cerebral cortex and mouse kidney tissues. ICC/IF: HeLa cells.
  • General notes

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

    ab227560 is a PBS only buffer version of ab40675, containing no BSA or sodium azide, ideal for antibody labeling. Please refer to ab40675 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab227560 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 at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 63 kDa).
ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

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, EP592Y, in a different buffer formulation (cat# ab40675).

    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 - ab40675 observed at 65 kDa. Red - loading control, ab8245, observed at 37 kDa.

     

    Ab40675 was shown to specifically react with TRAF6 in wild-type cells, along with additional cross-reactive bands as signal was lost in TRAF6 knockout HAP1 cells. Wild-type and TRAF6 knockout samples were subjected to SDS-PAGE. Ab40675 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue labelling TRAF6 with purified ab40675 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40675).

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TRAF6 with purified ab40675 at 1/50. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/50) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40675).

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TRAF6 with unpurified ab40675 at 1/250.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab40675).

  • This IHC data was generated using the same anti-TRAF6 antibody clone, EP592Y, in a different buffer formulation (cat# ab40675).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue labelling TRAF6 with purified ab40675 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

References

This product has been referenced in:
  • Huang H  et al. The effect of marrow stromal cells on TRAF6 expression levels in myeloma cells. Oncol Lett 14:1464-1470 (2017). Read more (PubMed: 28789366) »
  • Chen G  et al. Limb Remote Ischemic Postconditioning Reduces Ischemia-Reperfusion Injury by Inhibiting NADPH Oxidase Activation and MyD88-TRAF6-P38MAP-Kinase Pathway of Neutrophils. Int J Mol Sci 17: (2016). WB ; Rat . Read more (PubMed: 27898007) »
See all 2 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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