Recombinant Anti-TRAF6 antibody [EP592Y] - BSA and Azide free (ab227560)
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 - BSA and Azide free
- Suitable for: WB, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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 -
Specificity
This antibody is unsuitable for detecting tissue lysates in WB application.
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Tested applications
Suitable for: WB, ICC/IF, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, Daudi, Jurkat, HEK293 and HeLa cell lysates. IHC-P: Human cerebral cortex and mouse kidney tissues. ICC/IF: HeLa cells.
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General notes
ab227560 is the carrier-free version of ab40675.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
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.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP592Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 58 kDa (predicted molecular weight: 63 kDa).
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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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
modificationsSumoylated 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
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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
see all -
Alternative names
- E3 ubiquitin-protein ligase TRAF6 antibody
- Interleukin 1 signal transducer antibody
- Interleukin-1 signal transducer antibody
see all
Images
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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?This data was developed using the same antibody clone in a different buffer formulation (ab40675).
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.
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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).
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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).
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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.
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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.
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.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (11)
ab227560 has been referenced in 11 publications.
- Yang G & Zhao Y MicroRNA-490-3p inhibits inflammatory responses in LPS-induced acute lung injury of neonatal rats by suppressing the IRAK1/TRAF6 pathway. Exp Ther Med 21:152 (2021). PubMed: 33456519
- Zhao S et al. MicroRNA-602 prevents the development of inflammatory bowel diseases in a microbiota-dependent manner. Exp Ther Med 22:1373 (2021). PubMed: 34659519
- Yan K et al. A20 inhibits osteoclastogenesis via TRAF6-dependent autophagy in human periodontal ligament cells under hypoxia. Cell Prolif 53:e12778 (2020). PubMed: 32027437
- Wang X et al. Danshen (Salvia miltiorrhiza) restricts MD2/TLR4-MyD88 complex formation and signalling in acute myocardial infarction-induced heart failure. J Cell Mol Med 24:10677-10692 (2020). PubMed: 32757377
- Xue YL et al. Long non-coding RNA MEG3 inhibits M2 macrophage polarization by activating TRAF6 via microRNA-223 down-regulation in viral myocarditis. J Cell Mol Med 24:12341-12354 (2020). PubMed: 33047847