Name: Anti-TNFAIP3 antibody [EPR2663]
Brand: RabMAb
SKU: ab92324
Rating: 5 (1 reviews)

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Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR2663] to TNFAIP3
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Mouse, Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alkaline Phosphatase Carrier Free HRP

Product name

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

Rabbit monoclonal [EPR2663] to TNFAIP3
Host species

Rabbit
Specificity

Mouse species is recommended based on WB results, we do not guarantee IHC-P for mouse.
Tested applications

Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt (Intra) or ICC/IF
Species reactivity

Reacts with: Mouse, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: WEHI-3 treated with TNF (ab9642), Jurkat treated with TNF (ab9642) + TPA, Jurkat treated with 5ng/ml PMA for 48 hours and then treated with 2Âµg/ml PHA for 48 hours, HeLa, A549 and Daudi cell lysates. IHC-P: Human kidney tissue.
General notes
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
For more information see here.
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR2663
Isotype

IgG
Research areas
- Neuroscience
- Processes

Alternative Versions
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
KO cell lysates
Positive Controls
- Daudi whole cell lysate (ab3951)
Recombinant Protein
- Recombinant Human TNFAIP3 protein (ab132088)
Related Products
- Recombinant human TNF alpha protein (ab9642)

The Abpromise guarantee

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
WB		1/1000 - 1/5000. Predicted molecular weight: 90 kDa.
IHC-P	(1)	1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Mouse species is recommended based on WB results, we do not guarantee IHC-P for mouse.

Notes
WB 1/1000 - 1/5000. Predicted molecular weight: 90 kDa.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Mouse species is recommended based on WB results, we do not guarantee IHC-P for mouse.

Application notes

Is unsuitable for Flow Cyt (Intra) or ICC/IF.

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.
Target information above from: UniProt accession P21580 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 7128 Human
- Entrez Gene: 21929 Mouse
- Omim: 191163 Human
- SwissProt: P21580 Human
- SwissProt: Q60769 Mouse
- Unigene: 211600 Human
- Unigene: 116683 Mouse
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

Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324)

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

Lane 1 : Wild-type A549 cell lysate
Lane 2 : TNFAIP3 knockout A549 cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : TNFAIP3 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 90 kDa
Observed band size: 90 kDa

Lanes 1- 4: Merged signal (red and green). Green - ab92324 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab92324 was shown to react with TNFAIP3 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab266946 (knockout cell lysate ab257114) was used. Wild-type A549 and TNFAIP3 knockout A549 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. ab92324 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TNFAIP3 antibody [EPR2663] (ab92324)

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.
Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324)

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

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

Lanes 1-4: Merged signal (red and green). Green - ab92324 observed at 80 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab92324 Anti-TNFAIP3 antibody [EPR2663] 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. ab92324 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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 10000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324)

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

Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : TNFAIP3 knockout A549 (Human lung carcinoma cell line) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) 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 (Human T cell leukemia cell line from peripheral blood) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

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

Lanes 1-4: Merged signal (red and green). Green - ab92324 observed at 80 kDa. Red - loading control ab8245 observed at 36 kDa.

ab92324 Anti-TNFAIP3 antibody [EPR2663] was shown to specifically react with TNFAIP3 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266945 (knockout cell lysate ab257113) was used. Wild-type and TNFAIP3 knockout samples were subjected to SDS-PAGE. ab92324 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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.
Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324)

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?
Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324)

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.
Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324)

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

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.
Western blot - Anti-TNFAIP3 antibody [EPR2663] (ab92324)

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
Anti-TNFAIP3 antibody [EPR2663] (ab92324)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab92324? Please let us know so that we can cite the reference in this datasheet.

ab92324 has been referenced in 31 publications.

Li Y et al. Paired Box 5-Induced LINC00467 Upregulation Promotes the Progression of Laryngeal Squamous Cell Cancer by Triggering the MicroRNA-4735-3p/TNF Alpha-Induced Protein 3 Pathway. Mol Biotechnol 65:655-667 (2023). PubMed: 36214976
Geismann C et al. NF-κB/RelA controlled A20 limits TRAIL-induced apoptosis in pancreatic cancer. Cell Death Dis 14:3 (2023). PubMed: 36596765
Wang W et al. Fibroblast A20 governs fibrosis susceptibility and its repression by DREAM promotes fibrosis in multiple organs. Nat Commun 13:6358 (2022). PubMed: 36289219
Yang X et al. TRIM56 promotes malignant progression of glioblastoma by stabilizing cIAP1 protein. J Exp Clin Cancer Res 41:336 (2022). PubMed: 36471347
Mzyk P et al. A20 Attenuates the Fibrotic Response in the Trabecular Meshwork. Int J Mol Sci 23:N/A (2022). PubMed: 35216043

View all Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Breast tumor)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: TE pH9

Permeabilization

Specification

Breast tumor

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C

Fixative

10% normal buffered formalin

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.

Abcam user community

Verified customer

Submitted Dec 31 2019

Question

Dear Sirs,
Now, for our projects we
want to test an anti TNFAIP3 (A20) antibody on mouse brain
tissue. We known that Abcam produces some of them, but
neuther is specific and functioning on brain tissue. Can
we prove some of them? Can we receve some free aliquots?
We known that in vivo the functioning antibody are:
ab74037 Anti-TNFAIP3 antibody Rabbit polyclonal
ab92324 Anti-TNFAIP3 antibody [EPR2663] Rabbit monoclonal
ab111192 Anti-TNFAIP3 antibody Goat polyclonal
Looking forward to hear from you
Sincerely

Abcam community

Verified customer

Asked on Jul 03 2012

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.

Abcam Scientific Support

Answered on Jul 03 2012

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Positive Controls

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Domain

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (31)

Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-TNFAIP3 antibody [EPR2663]