重组Anti-TRAF6抗体[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
Related conjugates and formulations
概述
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产品名称
Anti-TRAF6抗体[EP592Y]
参阅全部 TRAF6 一抗 -
描述
兔单克隆抗体[EP592Y] to TRAF6 -
宿主
Rabbit -
特异性
This antibody is unsuitable for detecting tissue lysates in WB application.
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经测试应用
适用于: IHC-P, ICC/IF, WBmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- 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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常规说明
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EP592Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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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
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab40675于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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IHC-P |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/50.
For unpurified use at 1/250 - 1/500. |
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WB | (1) |
1/5000. Detects a band of approximately 58 kDa (predicted molecular weight: 63 kDa).
For unpurified use at 1/2000 - 1/10000. This antibody is unsuitable for detecting tissue lysates. |
说明 |
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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. This antibody is unsuitable for detecting tissue lysates. |
靶标
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功能
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. -
组织特异性
Expressed in heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. -
通路
Protein modification; protein ubiquitination. -
序列相似性
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. -
结构域
The coiled coil domain mediates homo- and hetero-oligomerization.
The MATH/TRAF domain binds to receptor cytoplasmic domains. -
翻译后修饰
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. -
细胞定位
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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数据库链接
- 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 -
别名
- E3 ubiquitin-protein ligase TRAF6 antibody
- Interleukin 1 signal transducer antibody
- Interleukin-1 signal transducer antibody
see all
图片
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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?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) - 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.
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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).
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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.
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling TRAF6 with unpurified ab40675 at 1/250.
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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.
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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.
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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? -
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 secondsThis antibody is unsuitable for detecting tissue lysates.
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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
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
文献 (42)
ab40675 被引用在 42 文献中.
- Jiang H et al. Acupuncture Ameliorates Depression-Like Behaviors Through Modulating the Neuroinflammation Mediated by TLR4 Signaling Pathway in Rats Exposed to Chronic Restraint Stress. Mol Neurobiol 61:2606-2619 (2024). PubMed: 37917302
- Wen S et al. MIR222HG attenuates macrophage M2 polarization and allergic inflammation in allergic rhinitis by targeting the miR146a-5p/TRAF6/NF-κB axis. Front Immunol 14:1168920 (2023). PubMed: 37205104
- Li ZL et al. Human osteoarthritic articular cartilage stem cells suppress osteoclasts and improve subchondral bone remodeling in experimental knee osteoarthritis partially by releasing TNFAIP3. Stem Cell Res Ther 14:253 (2023). PubMed: 37752608
- Huang J et al. Effect of curcumin on regulatory B cells in chronic colitis mice involving TLR/MyD88 signaling pathway. Phytother Res 37:731-742 (2023). PubMed: 36196887
- Liang XS et al. IRAK-M Ablation Promotes Status Epilepticus-Induced Neuroinflammation via Activating M1 Microglia and Impairing Excitatory Synaptic Function. Mol Neurobiol 60:5199-5213 (2023). PubMed: 37277682