Overview

  • Product name

    Anti-TNFAIP3 antibody [EPR2663]
    See all TNFAIP3 primary antibodies
  • Description

    Rabbit monoclonal [EPR2663] to TNFAIP3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human TNFAIP3 aa 500-600. The exact sequence is proprietary.
    (Peptide available as ab175807)

  • Positive control

    • WB: WEHI-3 treated with TNF (ab9642), Jurkat treated with TNF (ab9642) and TPA and Daudi cell lysates. IHC-P: Human kidney tissue. ICC/IF: Daudi and HeLa cells. Flow Cyt: HepG2 and Daudi cells.
  • General notes

    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.

    This product is a recombinant rabbit monoclonal antibody.

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 - 1/500.
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

  • Function

    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.
  • Sequence similarities

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

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

    Cytoplasm. Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • 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

Images

  • All lanes : Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/5000 dilution

    Lane 1 : WEHI-3 (Mouse leukemia lymphoblast) whole cell lysate
    Lane 2 : WEHI-3 treated with 20 ng/ml TNF alpha (ab9642) for 6 h

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 90 kDa
    Observed band size: 80 kDa
    why is the actual band size different from the predicted?

  • 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.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labelling TNFAIP with purified ab92324 at 1/500. Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/2000 dilution (unpurified) + Jurkat cell lysate - treated with TNF (ab9642) 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 and dilution buffer: 5% NFDM/TBST.

  • Anti-TNFAIP3 antibody [EPR2663] (ab92324) at 1/3000 dilution (purified) + Jurkat cell lysate - treated with TNF (ab9642) 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 and dilution buffer: 5% NFDM/TBST.

  • 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).

  • 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.

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

    Lane 1 : Jurkat cells treated with TNF (ab9642) and TPA
    Lane 2 : Daudi cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 90 kDa

References

This product has been referenced in:

  • Liu K  et al. Functional role of a long non-coding RNA LIFR-AS1/miR-29a/TNFAIP3 axis in colorectal cancer resistance to pohotodynamic therapy. Biochim Biophys Acta N/A:N/A (2018). WB . Read more (PubMed: 29807108) »
  • Kwon DJ  et al. Generation of a-1,3-galactosyltransferase knocked-out transgenic cloned pigs with knocked-in five human genes. Transgenic Res 26:153-163 (2017). WB, IHC-P . Read more (PubMed: 27554374) »
See all 12 Publications for this product

Customer reviews and Q&As

Answer

Thank you for contacting us.

Because we carry over 90,000 products, it isn't feasible for us to keep small sample sizes of our products, and therefore I am not able to offer a test sample of any of the antibodies you are interested in.

However, ab74037 has been validated with mouse samples by IHC (in frozen sections), and therefore it is guaranteed to work in this application, so if it did not work following the datasheet indications, I would be happy to send a replacement or reimbursement as stated by our Abpromise guarantee.

Unfortunately ab92324 has been proved not to react with mouse samples, so I would not recommend using this antibody for your purpose. On the other hand ab111192 has only been validated to work with human samples in IHC (paraffin embedded sections).

My recommendation would be using ab74037 with mouse tissue if you are using IHC-Fr. If you wish to use it for IHC-P I would be happy to offer a discount code to test it in this technique and submit your feedback to us.

Therefore, I can offer a discount off a future purchase if you buy ab74037 now, test it in IHC-P and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please reply to me letting me know before purchasing the antibody.

To find out more about our Abreview system, please visit: https://www.abcam.com/abreviews.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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