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Rabbit Recombinant Monoclonal TRAF6 antibody. Suitable for IHC-P, WB, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 152 publications.


Images

Western blot - Anti-TRAF6 antibody [EP591Y] (AB33915), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-TRAF6 antibody [EP591Y] (AB33915), expandable thumbnail
  • Western blot - Anti-TRAF6 antibody [EP591Y] (AB33915), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-TRAF6 antibody [EP591Y] (AB33915), expandable thumbnail
  • Western blot - Anti-TRAF6 antibody [EP591Y] (AB33915), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBFlow Cyt (Intra)
Human
Tested
Not recommended
Tested
Tested
Mouse
Expected
Not recommended
Tested
Expected
Rat
Expected
Not recommended
Tested
Expected
Zebrafish
Predicted
Not recommended
Predicted
Predicted

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Zebrafish

Dilution info

-

Notes

-

Not recommended
Not recommended

Species

Mouse, Rat, Human, Zebrafish

Dilution info

-

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/2000 - 1/10000

Notes

-

Species

Rat

Dilution info

1/2000 - 1/10000

Notes

-

Species

Human

Dilution info

1/2000 - 1/10000

Notes

-

Predicted
Predicted

Species

Zebrafish

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/50

Notes

Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Predicted
Predicted

Species

Zebrafish

Dilution info

-

Notes

-

Associated Products

Select an associated product type

5 products for Alternative Product

Target data

Function

E3 ubiquitin ligase that, together with UBE2N and UBE2V1, mediates the synthesis of 'Lys-63'-linked-polyubiquitin chains conjugated to proteins, such as ECSIT, IKBKG, IRAK1, AKT1 and AKT2 (PubMed:11057907, PubMed:18347055, PubMed:19465916, PubMed:19713527, PubMed:31620128). Also mediates ubiquitination of free/unanchored polyubiquitin chain that leads to MAP3K7 activation (PubMed:19675569). Leads to the activation of NF-kappa-B and JUN (PubMed:16378096, PubMed:17135271, PubMed:17703191). Seems to also play a role in dendritic cells (DCs) maturation and/or activation (By similarity). Represses c-Myb-mediated transactivation, in B-lymphocytes (PubMed:18093978, PubMed:18758450). Adapter protein that seems to play a role in signal transduction initiated via TNF receptor, IL-1 receptor and IL-17 receptor (PubMed:12140561, PubMed:19825828, PubMed:8837778). Regulates osteoclast differentiation by mediating the activation of adapter protein complex 1 (AP-1) and NF-kappa-B, in response to RANK-L stimulation (By similarity). Together with MAP3K8, mediates CD40 signals that activate ERK in B-cells and macrophages, and thus may play a role in the regulation of immunoglobulin production (By similarity). Participates also in the TCR signaling by ubiquitinating LAT (PubMed:23514740, PubMed:25907557).

Alternative names

RNF85, RNF85, TRAF6, TNF receptor-associated factor 6, E3 ubiquitin-protein ligase TRAF6, Interleukin-1 signal transducer, RING finger protein 85, RING-type E3 ubiquitin transferase TRAF6

Recommended products

Rabbit Recombinant Monoclonal TRAF6 antibody. Suitable for IHC-P, WB, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 152 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EP591Y

Purification technique

Affinity purification Protein A

Specificity

This antibody is unsuitable for detecting TRAF6 in tissue lysates.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

TRAF6 also known as TNF receptor-associated factor 6 is an E3 ubiquitin ligase that is part of the TRAF family. It has a molecular mass of approximately 60 kDa. TRAF6 is ubiquitously expressed in many types of tissues and cells including immune cells like macrophages and dendritic cells. Its primary role is to mediate signal transduction from members of the TNF receptor and Toll-like receptor (TLR) families which it achieves through polyubiquitination of target proteins. This process facilitates the recruitment of downstream signaling components necessary for cellular responses.

Biological function summary

TRAF6 plays a critical role in regulating immune and inflammatory responses. It acts within a complex often associating with other signaling proteins such as IRAK1 and TAK1 to activate downstream kinases like IKK and JNK. These signaling pathways lead to the activation of transcription factors such as NF-kB and AP-1 which regulate the expression of genes involved in inflammation and immune responses. TRAF6 is an important modulator of adaptive and innate immunity influencing both the maturation of immune cells and the body's response to infection.

Pathways

TRAF6 significantly contributes to the NF-kB and MAPK signaling pathways. In the NF-kB pathway TRAF6 mediates signal transduction following TLR engagement interacting with proteins such as MyD88 and RIP1 to drive inflammatory cytokine production. In the MAPK pathway it associates predominantly with TAK1 to facilitate the phosphorylation and activation of downstream kinases. These pathways are essential for transmitting signals from cell surface receptors to the nucleus controlling the magnitude of immune signaling and cellular stress responses.

Associated diseases and disorders

TRAF6 has a significant association with autoimmune diseases and certain cancers. In autoimmune disorders aberrant activation of TRAF6 can lead to prolonged inflammation and detrimental immune responses. For cancers TRAF6 is linked to tumor progression and metastasis acting alongside proteins like IRAK1 and RIP1 which can further promote malignancy through chronic inflammation and immune evasion. Understanding TRAF6's role in these diseases provides insights that could lead to targeted therapeutic strategies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

9 product images

  • Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915), expandable thumbnail

    Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915)

    Lanes 1- 4: Merged signal (red and green). Green - ab33915 observed at 65 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

    ab33915 was shown to react with TRAF6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human TRAF6 knockout HeLa cell line ab266009 (knockout cell lysate Human TRAF6 knockout HeLa cell lysate ab257760) was used. Wild-type HeLa and TRAF6 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab33915 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915) at 1/2000 dilution

    Lane 1: Wild-type HeLa cell lysate at 20 µg

    Lane 2: TRAF6 knockout HeLa cell lysate at 20 µg

    Lane 3: HAP1 cell lysate at 20 µg

    Lane 4: Daudi cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 60 kDa

    Observed band size: 65 kDa

  • Flow Cytometry (Intracellular) - Anti-TRAF6 antibody [EP591Y] (ab33915), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-TRAF6 antibody [EP591Y] (ab33915)

    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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG isotype control antibody (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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.

  • Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915), expandable thumbnail

    Western blot - Anti-TRAF6 antibody [EP591Y] (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, Anti-GAPDH antibody [6C5] - 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 Anti-GAPDH antibody [6C5] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging

    All lanes: Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915)

    Predicted band size: 60 kDa

  • Flow Cytometry (Intracellular) - Anti-TRAF6 antibody [EP591Y] (ab33915), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-TRAF6 antibody [EP591Y] (ab33915)

    Intracellular 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.

  • Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915), expandable thumbnail

    Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915)

    All lanes: Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915) at 1/10000 dilution

    All lanes: Jurkat cell lysate

    Predicted band size: 60 kDa

  • Flow Cytometry (Intracellular) - Anti-TRAF6 antibody [EP591Y] (ab33915), expandable thumbnail
    This image is courtesy of an anonymous customer review.

    Flow Cytometry (Intracellular) - Anti-TRAF6 antibody [EP591Y] (ab33915)

    ab33915 staining TRAF6 in Human platelet cells by intracellular 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)

  • Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915), expandable thumbnail

    Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915)

    All lanes: Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915) at 1/1000 dilution

    Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

    Lane 2: Human heart lysates at 15 µg

    Lane 3: Human skeletal muscle lysates at 15 µg

    Lane 4: Mouse skeletal muscle lysates at 15 µg

    Lane 5: Rat skeletal muscle lysates at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 60 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP591Y] (ab33915), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP591Y] (ab33915)

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

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915), expandable thumbnail

    Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915)

    All lanes: Western blot - Anti-TRAF6 antibody [EP591Y] (ab33915) at 1/1000 dilution

    Lane 1: Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 15 µg

    Lane 2: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 15 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 60 kDa

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Product protocols

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