Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-TRAF6 antibody [EP591Y] (Alexa Fluor® 488) (ab214638)

Overview

  • Product name

    Anti-TRAF6 antibody [EP591Y] (Alexa Fluor® 488)
    See all TRAF6 primary antibodies
  • Description

    Rabbit monoclonal [EP591Y] to TRAF6 (Alexa Fluor® 488)
  • Host species

    Rabbit
  • Conjugation

    Alexa Fluor® 488. Ex: 495nm, Em: 519nm
  • Tested applications

    Suitable for: Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human TRAF6 aa 50-150. The exact sequence is proprietary.
    Database link: Q9Y4K3
    (Peptide available as ab183540)

  • Positive control

    • ICC/IF: HeLa cells Flow Cyt: HAP1-wt cells.
  • General notes

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

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab214638 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 a concentration of 0.1 µg/ml.
ICC/IF 1/100.

This product gave a positive signal in HeLa cells fixed with 100% methanol (5 min)

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

  • Overlay histogram showing HAP1 wildtype (green line) and HAP1-TRAF6 knockout cells (red line) stained with ab214638. 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 (ab214638, 0.1µg/ml) for 30 min at 22°C.

    A rabbit IgG isotype control antibody  (ab199091) 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.

  • ab214638 staining TRAF6 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab214638 at 1/100 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

References

ab214638 has not yet been referenced specifically in any publications.

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