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    traf6-antibody-ep592y-ab40675.pdf

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Cell Biology Apoptosis Receptors Associated Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-TRAF6 antibody [EP592Y] (ab40675)

  • Datasheet
  • SDS
Reviews (1)Q&A (1)References (18)

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Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP592Y] (ab40675)
  • Immunocytochemistry/ Immunofluorescence - Anti-TRAF6 antibody [EP592Y] (ab40675)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP592Y] (ab40675)
  • Immunocytochemistry/ Immunofluorescence - Anti-TRAF6 antibody [EP592Y] (ab40675)
  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
  • Anti-TRAF6 antibody [EP592Y] (ab40675)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP592Y] to TRAF6
  • Suitable for: IHC-P, ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Secondary
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Knockout
Product image
Human TRAF6 knockout HeLa cell line (ab266009)

View more associated products

Overview

  • Product name

    Anti-TRAF6 antibody [EP592Y]
    See all TRAF6 primary antibodies
  • Description

    Rabbit monoclonal [EP592Y] to TRAF6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Purchase matching WB positive control:Recombinant Human TRAF6 protein
    • WB: HAP1, Daudi, Jurkat, HEK293 and HeLa cell lysates. IHC-P: Human cerebral cortex and mouse kidney tissues. ICC/IF: HeLa cells. This antibody is unsuitable for detecting TRAF6 in tissue lysates.
  • General notes

    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 production
    For more information see here.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP592Y
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Receptors
    • Associated Proteins
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • NFkB Pathway
    • Signal Transduction
    • Growth Factors/Hormones
    • TNF
    • Microbiology
    • Organism
    • Virus
    • RNA Virus
    • ssRNA positive strand virus
    • SARS Coronavirus
    • Cancer
    • Growth factors
    • TNF
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Death receptors & ligands
    • TRAF
    • Immunology
    • Innate Immunity
    • TLR Signaling
    • Cancer
    • Cell Death
    • Apoptosis
    • Receptors
    • Death receptors & ligands
    • TRAF
    • Cancer
    • Cell Death
    • Apoptosis
    • Receptors
    • Associated Proteins

Associated products

  • Alternative Versions

    • Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human TRAF6 knockout HeLa cell line (ab266009)
  • KO cell lysates

    • Human TRAF6 knockout HeLa cell lysate (ab257760)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab30128)
    • Jurkat whole cell lysate (ab7899)
    • HEK-293 whole cell lysate (ab7902)
  • Recombinant Protein

    • Recombinant Human TRAF6 protein (ab132023)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab40675 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
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/50.

For unpurified use at 1/250 - 1/500.

WB (1)
1/5000. Detects a band of approximately 58 kDa (predicted molecular weight: 63 kDa).

For unpurified use at 1/2000 - 1/10000.

Notes
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/50.

For unpurified use at 1/250 - 1/500.

WB
1/5000. Detects a band of approximately 58 kDa (predicted molecular weight: 63 kDa).

For unpurified use at 1/2000 - 1/10000.

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.
  • Target information above from: UniProt accession Q9Y4K3 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7189 Human
    • Entrez Gene: 22034 Mouse
    • Entrez Gene: 311245 Rat
    • Omim: 602355 Human
    • SwissProt: Q9Y4K3 Human
    • SwissProt: P70196 Mouse
    • SwissProt: B5DF45 Rat
    • Unigene: 591983 Human
    • Unigene: 292729 Mouse
    • Unigene: 220435 Rat
    see all
  • 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

  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    All lanes : Anti-TRAF6 antibody [EP592Y] (ab40675) at 1/500 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : TRAF6 knockout HeLa cell lysate
    Lane 3 : HAP1 cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 63 kDa
    Observed band size: 65 kDa why is the actual band size different from the predicted?



    Lanes 1- 4: Merged signal (red and green). Green - ab40675 observed at 65 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab40675 was shown to react with TRAF6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab266009 (knockout 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. ab40675 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP592Y] (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.

  • Immunocytochemistry/ Immunofluorescence - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Immunocytochemistry/ Immunofluorescence - Anti-TRAF6 antibody [EP592Y] (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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRAF6 antibody [EP592Y] (ab40675)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Immunocytochemistry/ Immunofluorescence - Anti-TRAF6 antibody [EP592Y] (ab40675)

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

  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    All lanes : Anti-TRAF6 antibody [EP592Y] (ab40675) at 1/5000 dilution (purified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 63 kDa
    Observed band size: 58 kDa why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Anti-TRAF6 antibody [EP592Y] (ab40675) at 1/1000 dilution (purified) + HEK293 cell lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 63 kDa
    Observed band size: 58 kDa why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

     

  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Anti-TRAF6 antibody [EP592Y] (ab40675) at 1/2000 dilution (unpurified) + 10ug Jurkat cell lysate

    Predicted band size: 63 kDa
    Observed band size: 58 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    All lanes : Anti-TRAF6 antibody [EP592Y] (ab40675) at 1/1000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : Human heart lysates
    Lane 3 : Human skeletal muscle lysates
    Lane 4 : Mouse skeletal muscle lysates
    Lane 5 : Rat skeletal muscle lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 63 kDa


    Exposure time: 70 seconds


    This antibody is unsuitable for detecting tissue lysates.  

  • Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    Western blot - Anti-TRAF6 antibody [EP592Y] (ab40675)
    All lanes : Anti-TRAF6 antibody [EP592Y] (ab40675) at 1/1000 dilution

    Lane 1 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 63 kDa

  • Anti-TRAF6 antibody [EP592Y] (ab40675)
    Anti-TRAF6 antibody [EP592Y] (ab40675)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (18)

Publishing research using ab40675? Please let us know so that we can cite the reference in this datasheet.

ab40675 has been referenced in 18 publications.

  • Zhao L  et al. The Pharmacological Effect and Mechanism of Lanthanum Hydroxide on Vascular Calcification Caused by Chronic Renal Failure Hyperphosphatemia. Front Cell Dev Biol 9:639127 (2021). PubMed: 33928079
  • Liu T  et al. Genome-wide characterization of TNF receptor-associated factors in the Chinese soft-shelled turtle Pelodiscus sinensis and their expression profiling in response to Aeromonas hydrophila challenge. Fish Shellfish Immunol 101:88-98 (2020). PubMed: 32229294
  • Gao J  et al. Bone marrow mesenchymal stem cells improve bone erosion in collagen-induced arthritis by inhibiting osteoclasia-related factors and differentiating into chondrocytes. Stem Cell Res Ther 11:171 (2020). PubMed: 32381074
  • Paparo L  et al. Protective action of Bacillus clausii probiotic strains in an in vitro model of Rotavirus infection. Sci Rep 10:12636 (2020). PubMed: 32724066
  • Zhang DH  et al. Deubiquitinase Ubiquitin-Specific Protease 10 Deficiency Regulates Sirt6 signaling and Exacerbates Cardiac Hypertrophy. J Am Heart Assoc 9:e017751 (2020). PubMed: 33170082
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-2 of 2 Abreviews or Q&A

Western blot abreview for Anti-TRAF6 antibody [EP592Y]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (mouse lung)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
reperfusion
Specification
mouse lung
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 21 2018

Question

I had a short conversation this morning with Kasia from livesupport about a TRAF6 monoclonal antibody for studies in zebrafish. As the antibodies (clone EP591Y and EP592Y) that you are offering are not yet tested in zebrafish I would like to ask you to check if the sequence of the synthetic peptide is conserved in zebrafish. I attached a text file with an alignment of mouse, human and zebrafish TRAF6. Kind regards,

Read More

Abcam community

Verified customer

Asked on Jul 24 2007

Answer

Thank you for your enquiry. I am pleased to inform you that after consulting the source of these two antibodies we have the following information: With ab33915 its epitope shows 100% aa homology to zebra fish sequence. While with ab40675, its epitope showed only 57% homology. If you need further information please feel free to contact me any time.

Read More

Abcam Scientific Support

Answered on Jul 24 2007

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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