Overview

  • Product nameAnti-TNFAIP3 antibody [EPR2663]
    See all TNFAIP3 primary antibodies
  • Description
    Rabbit monoclonal [EPR2663] to TNFAIP3
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human TNFAIP3 aa 500-600.
    (Peptide available as ab175807)

  • Positive control
    • WB: Jurkat cells treated with TNF and TPA and Daudi cell lysates. IHC-P: Human kidney tissues. ICC/IF: Daudi and HeLa cells. Flow Cyt: HepG2 and Daudi cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

    Alternative versions available:

    Anti-TNFAIP3 antibody (Alexa Fluor® 488) [EPR2663] (ab197541)
    Anti-TNFAIP3 antibody (Alexa Fluor® 647) [EPR2663] (ab197068)

Properties

Applications

Our Abpromise guarantee covers the use of ab92324 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
ICC/IF 1/50 - 1/100.
WB 1/1000 - 1/5000. Predicted molecular weight: 90 kDa.Can be blocked with TNFAIP3 peptide (ab175807).
IHC-P 1/50 - 1/100.
Flow Cyt 1/20 - 1/50. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • FunctionUbiquitin-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.
  • Sequence similaritiesBelongs to the peptidase C64 family.
    Contains 7 A20-type zinc fingers.
    Contains 1 OTU domain.
  • DomainThe A20-type zinc fingers mediate the ubiquitin ligase activity.
    The OTU domain mediates the deubiquitinase activity.
  • Cellular localizationCytoplasm. Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • A20 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

Anti-TNFAIP3 antibody [EPR2663] images

  • Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/2000 dilution (unpurified) + Jurkat cell lysate - treated with TNF and TPA at 10 µg

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

    Predicted band size : 90 kDa
    Observed band size : 80 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.

  • Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/3000 dilution (purified) + Jurkat cell lysate - treated with TNF and TPA at 10 µg

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

    Predicted band size : 90 kDa
    Observed band size : 80 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.

  • All lanes : Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/1000 dilution (unpurified)

    Lane 1 : Jurkat cells treated with TNF and TPA
    Lane 2 : Daudi cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP labelled goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size : 90 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling TNFAIP3 with unpurified ab92324 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling TNFAIP3 with purified ab92324 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

  • Immunocytochemsitry/Immunofluorescence analysis of Daudi cells labelling TNFAIP3 (red) with unpurified ab92324 at 1/50. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

  • Immunocytochemsitry/Immunofluorescence analysis of Daudi cells labelling TNFAIP3 (red) with purified ab92324 at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

  • ICC/IF image of unpurified ab92324 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab92324, neat) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HepG2 cells stained with unpurified ab92324 (red line). The cells were fixed with 80% 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. The cells were then incubated with the antibody (ab92324, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Flow cytometry analysis of Daudi cells labelling TNFAIP3 with unpurified ab92324 at 1/20 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

  • Flow cytometry analysis of Daudi cells labelling TNFAIP3 with purified ab92324 at 1/30 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

References for Anti-TNFAIP3 antibody [EPR2663] (ab92324)

This product has been referenced in:
  • Troppan K  et al. Frequent down regulation of the tumor suppressor gene a20 in multiple myeloma. PLoS One 10:e0123922 (2015). Read more (PubMed: 25856582) »
  • de la Rica L  et al. NF-?B-direct activation of microRNAs with repressive effects on monocyte-specific genes is critical for osteoclast differentiation. Genome Biol 16:2 (2015). WB . Read more (PubMed: 25601191) »

See all 5 Publications for this product

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