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使用敲除细胞株进行验证

Anti-TNFAIP3抗体[59A426] (ab13597)

Reviews (1)Q&A (2)References (39)
Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597)
  • Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNFAIP3 antibody [59A426] (ab13597)
  • Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597)
  • Flow Cytometry (Intracellular) - Anti-TNFAIP3 antibody [59A426] (ab13597)

Key features and details

  • Mouse monoclonal [59A426] to TNFAIP3
  • Suitable for: Flow Cyt (Intra), IHC-P, WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

选择批间可重复性更高的重组抗体

Product image
Anti-TNFAIP3 antibody [EPR2663] (ab92324)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术，可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-TNFAIP3抗体[59A426]
    参阅全部 TNFAIP3 一抗

  • 描述

    小鼠单克隆抗体[59A426] to TNFAIP3

  • 宿主

    Mouse

  • 经测试应用

    适用于: Flow Cyt (Intra), IHC-P, WBmore details
    不适用于: ICC

  • 种属反应性

    与反应: Human

  • 免疫原

    Recombinant full length protein corresponding to Human TNFAIP3.
    Database link: P21580

  • 表位

    The epitope has been mapped to the C-terminal portion of A20, amino acids 440-790.

  • 阳性对照

    • WB: Daudi and HeLa cell lysates. Flow Cyt (Intra): HepG2 cells

  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab13597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
Flow Cyt (Intra)
Use 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P
Use at an assay dependent concentration.
WB (1)
Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 70 kDa.
说明
Flow Cyt (Intra)
Use 1-2µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-P
Use at an assay dependent concentration.
WB
Use a concentration of 2 - 4 µg/ml. Detects a band of approximately 70 kDa.
应用说明
Is unsuitable for ICC.

靶标

  • 功能

    Ubiquitin-editing enzyme that contains both ubiquitin ligase and deubiquitinase activities. Essential component of a ubiquitin-editing protein complex, comprising also RNF11, ITCH and TAX1BP1, that ensures the transient nature of inflammatory signaling pathways. Upon TNF stimulation, deubiquitinates 'Lys-63'-polyubiquitin chains on RIPK1 and catalyzes the formation of 'Lys-48'-polyubiquitin chains. This leads to RIPK1 proteasomal degradation and consequently termination of the TNF- or LPS-mediated activation of NF-kappa-B. In vitro able to deubiquitinate both 'Lys-48'- and 'Lys-63' polyubiquitin chains. Inhibitor of programmed cell death. Has a role in the function of the lymphoid system.

  • 序列相似性

    Belongs to the peptidase C64 family.
    Contains 7 A20-type zinc fingers.
    Contains 1 OTU domain.

  • 结构域

    The A20-type zinc fingers mediate the ubiquitin ligase activity.
    The OTU domain mediates the deubiquitinase activity.

  • 细胞定位

    Cytoplasm. Nucleus.
  • Information by UniProt

  • 数据库链接

    • 别名

      • A20 antibody
      • AISBL antibody
      • MGC104522 antibody
      • MGC138687 antibody
      • MGC138688 antibody
      • OTU domain containing protein 7C antibody
      • OTU domain-containing protein 7C antibody
      • OTUD7C antibody
      • Putative DNA binding protein A20 antibody
      • Putative DNA-binding protein A20 antibody
      • TNAP3_HUMAN antibody
      • TNF alpha-induced protein 3 antibody
      • TNFA1P2 antibody
      • TNFAIP 3 antibody
      • TNFAIP3 (A20) antibody
      • TNFAIP3 antibody
      • Tumor necrosis factor alpha induced protein 3 antibody
      • Tumor necrosis factor alpha-induced protein 3 antibody
      • Tumor necrosis factor induced protein 3 antibody
      • Tumor necrosis factor inducible protein A20 antibody
      • tumor necrosis factor, alpha-induced protein 3 antibody
      • Zinc finger protein A20 antibody
      see all

    图片

    • Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597)
      Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597)
      All lanes : Anti-TNFAIP3 antibody [59A426] (ab13597) at 1/500 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : TNFAIP3 knockout HeLa cell lysate
      Lane 3 : Jurkat cell treated with 5ng/ml PMA for 48 hours and then treated with 2µg/ml PHA for 48 hours, whole cell lysate
      Lane 4 : Untreated Jurkat cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Observed band size: 80 kDa why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab13597 observed at 80 kDa. Red - loading control, ab181602 observed at 37 kDa.

      ab13597 Anti-TNFAIP3 antibody [59A426] was shown to specifically react with TNFAIP3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265983 (knockout cell lysate ab257112) was used. Wild-type and TNFAIP3 knockout samples were subjected to SDS-PAGE. ab13597 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

       

    • Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597)
      Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597)
      Anti-TNFAIP3 antibody [59A426] (ab13597) at 4 µg/ml + Jurkat cell lysate
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNFAIP3 antibody [59A426] (ab13597)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNFAIP3 antibody [59A426] (ab13597)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human placenta tissue labelling TNFAIP3 with ab13597 at 5µg/ml. Staining was enhanced by boiling tissue sections in 10mM sodium citrate buffer, pH6.0 for 10-20 minutes followed by cooling at room temperature for 20 minutes.

    • Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597)
      Western blot - Anti-TNFAIP3 antibody [59A426] (ab13597)
      All lanes : Anti-TNFAIP3 antibody [59A426] (ab13597) at 1 µg/ml

      Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
      Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Observed band size: 90 kDa why is the actual band size different from the predicted?
      Additional bands at: 15 kDa, 34 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 20 minutes
    • Flow Cytometry (Intracellular) - Anti-TNFAIP3 antibody [59A426] (ab13597)
      Flow Cytometry (Intracellular) - Anti-TNFAIP3 antibody [59A426] (ab13597)
      Overlay histogram showing HepG2 cells stained with ab13597 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab13597, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HepG2 cells fixed with methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    文献 (39)

    发表研究结果有使用 ab13597？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab13597 被引用在 39 文献中.

    • Xu X  et al. Decreased ubiquitin modifying enzyme A20 associated with hyper-responsiveness to ovalbumin challenge following intrauterine growth restriction. Respir Res 24:50 (2023). PubMed: 36788604
    • Rohwedder I  et al. A20 and the noncanonical NF-κB pathway are key regulators of neutrophil recruitment during fetal ontogeny. JCI Insight 8:N/A (2023). PubMed: 36633909
    • Fan L  et al. Effects of Ruanmailing in Blocking Early Stages of Atherosclerosis by TNF-α Regulation via Kir2.1. Evid Based Complement Alternat Med 2022:2836880 (2022). PubMed: 35432558
    • Deng HJ  et al. A20 Establishes Negative Feedback With TRAF6/NF-κB and Attenuates Early Brain Injury After Experimental Subarachnoid Hemorrhage. Front Immunol 12:623256 (2021). PubMed: 34381441
    • Ning B  et al. MSK1 downregulation is involved in inflammatory responses following subarachnoid hemorrhage in rats. Exp Ther Med 21:364 (2021). PubMed: 33732337
    View all Publications for this product

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    1-3 of 3 Abreviews or Q&A

    Western blot abreview for Anti-TNFAIP3 antibody [59A426]

     Average
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HCT116, HT29 and Caco2 colon cancer cell lines)
    Loading amount
    30 µg
    Specification
    HCT116, HT29 and Caco2 colon cancer cell lines
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% in TBST
    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.

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    Verified customer

    提交于 Nov 07 2006

    Question

    I saw as well this band around 85-89 and other 75 kda. After cleavage tnfiap3 full leigh two fragments are released, one with 37 kDa and other with 50 kDa. Some papers said that youreally antibody can recognize the 85-89 kDa band, 75 kDa band and the 37 kDa fragment, but not the 50 kDa.
    However I did identify the 50 kDa in my data, so do you have any data supporting this?

    Read More
    Answer

    There is data to suggest that there is a 58kD band present in many of the samples tested with this antibody. This was tested externally so I am sorry I am not able to provide the data itself. Analysis of A20 using this antibody in WB showed as a stack of bands and LPS treatment (200 ng) induced a ˜58 kDa putative A20 cleavage band  not seen in the untreated control (C) THP1 cells. A20 cleavage fragments have often been described to accompany NF-kB activation. Although the significance of the cleavage fragments remains to be fully elucidated, they are interesting because cleavage fragments often have key roles in signaling pathways.

    .

    Read More

    Question

    Will your mouse anti-A20 react with mouse A20 in Western blots?

    Read More
    Answer

    Thank you for your enquiry. Our two A20 antibodies raised in mice have been tested for Western blotting on human samples. Human A20 is 89% homologous with the sequence for mouse A20. So there is a good chance these antibodies will recognize mouse A20 but we cannot guarantee this. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

    Read More

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