A20/TNFAIP3 Antibody (59A426)

Images

 
Western Blot: A20/TNFAIP3 Antibody (59A426) [NBP1-77533] - Human Jurkat lysate probed with A20 antibody at 4 ug/ml.
Flow Cytometry: A20/TNFAIP3 Antibody (59A426) [NBP1-77533] - An intracellular stain was performed on SK-MEL-28 cells with A20/TNFAIP3 (59A426) antibody NBP1-77533AF647 (blue) and a matched isotype control (orange). ...read more
Simple Western: A20/TNFAIP3 Antibody (59A426) [NBP1-77533] - Lane view shows a specific band for A20/TNFAIP3 in 0.05 mg/ml of Jurkat lysate. This experiment was performed under reducing conditions using the 12-230 kDa ...read more
Immunohistochemistry-Paraffin: A20/TNFAIP3 Antibody (59A426) [NBP1-77533] - Human placenta probed with A20 antibody at 5 ug/ml, cytoplasmic staining of decidual cells is observed. Human tissue TMA was used for this ...read more
Immunocytochemistry/ Immunofluorescence: A20/TNFAIP3 Antibody (59A426) [NBP1-77533] - HepG2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The ...read more
Immunocytochemistry/ Immunofluorescence: A20/TNFAIP3 Antibody (59A426) [NBP1-77533] - Caco-2 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells ...read more
Flow Cytometry: A20/TNFAIP3 Antibody (59A426) [NBP1-77533] - An intracellular stain was performed on RH-30 cells with A20/TNFAIP3 Antibody (59A426) NBP1-77533PE (blue) and a matched isotype control (orange). Cells were ...read more

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications WB, Simple Western, Flow, Flow-IC, ICC/IF, IHC, IHC-P, IP, CyTOF-ready
Clone
59A426
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1.0 mg/ml

Order Details

A20/TNFAIP3 Antibody (59A426) Summary

Immunogen
Full length recombinant human A20/TNFAIP3 Antibody (59A426). The epitope has been mapped to the C-terminal portion of A20 (amino acids 440-790)
Isotype
IgG1 Kappa
Clonality
Monoclonal
Host
Mouse
Gene
TNFAIP3
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 2-4 ug/ml
  • Simple Western 1:100
  • Flow Cytometry 2.5 ug/ml
  • Immunocytochemistry/Immunofluorescence 10 ug/ml
  • Immunohistochemistry 5 - 10 ug/ml
  • Immunohistochemistry-Paraffin 5 - 10 ug/ml
  • Immunoprecipitation 1-2 ug/ml
  • CyTOF-ready
  • Flow (Intracellular) 2.5 ug/ml
Application Notes
The A20 antibody (clone 59A426) has also been used in IHC-P (Metellus, 2010) and Flow Cytometry (intracellular) (Hjelmeland, 2010). Multiple A20 cleavage fragments have been described in Western Blot, see Coornaert (2008) and Hailfinger (2009) for additional details.

In Simple Western only 10 - 15 ul of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. This antibody is CyTOF ready.
Control
A20/TNFAIP3 Knockout HeLa Cell Lysate
Publications
Read Publications using
NBP1-77533 in the following applications:

  • WB
    6 publications

Packaging, Storage & Formulations

Storage
Store at -20C. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

Alternate Names for A20/TNFAIP3 Antibody (59A426)

  • A20
  • A20TNFA1P2
  • EC 3.4.19.12
  • EC 6.3.2.-
  • MGC104522
  • MGC138687
  • OTU domain-containing protein 7C
  • OTUD7C
  • OTUD7CMGC138688
  • Putative DNA-binding protein A20
  • TNF alpha-induced protein 3
  • TNFA1P2
  • TNFAIP3
  • tumor necrosis factor alpha-induced protein 3
  • tumor necrosis factor inducible protein A20
  • tumor necrosis factor, alpha-induced protein 3
  • Zinc finger protein A20

Background

A20, TNFalpha-induced protein 3, TNFAIP3 (theoretical molecular weight 95 kDa) is an important regulator of pro-inflammatory signaling pathways such as NF-kB activation and tumor necrosis factor (TNF)-mediated programmed cell death (1, 2). In macrophages, A20/TNFAIP3 controls the release of IL-1 beta/IL-18 by regulating NLRP3 inflammasome activity and CXCL9/CXCL10 production via STAT1 signaling. By regulating the expression of inflammatory molecules such as IL-6 and anti-apoptotic proteins in dendritic cells, A20/TNFAIP3 maintains T-cell and B-cell homeostasis.

A20/TNFAIP3 is a highly conserved protein sharing >90% amino acid sequence identity across various mammalian species and is highly expressed in T-cell and B-cells. Coding and non-coding single nucleotide polymorphisms (SNPs) of A20/TNFAIP3 have been associated with multiple autoinflammatory and autoimmune diseases including type 1 diabetes, rheumatoid arthritis, Crohn's disease, and systemic lupus erythematosus (SLE) (3). The two domains of A20/TNFAIP3 cooperate to regulate NF-kB signaling. Its N-terminal ovarian tumor (OTU) domain contains a catalytic cysteine (C103) which functions as a K63 deubiquitinase, whereas the 7 zinc fingers that make up the C-terminal domain mediate K48 polyubiquitination. To regulate NF-kB signaling, A20/TNFAIP3 removes K63-polyubiquitin chains from receptor-interacting protein 1 (RIP1) and NF-kB essential modulator (NEMO), thus preventing interactions with downstream partners. A20/TNFAIP3 also contributes to the degradation of RIP1 and Ubc13 through the addition of K48 polyubiquitin chains (4).

References

1.Dixit VM1, Green S, Sarma V, Holzman LB, Wolf FW, O'Rourke K, Ward PA, Prochownik EV, Marks RM. (1990) Tumor necrosis factor-alpha induction of novel gene products in human endothelial cells including a macrophage-specific chemotaxin. J Biol Chem. 265(5):2973-8. PMID: 2406243

2.Verstrepen L, Verhelst K, van Loo G, Carpentier I, Ley SC, Beyaert R. (2010) Expression, biological activities and mechanisms of action of A20 (TNFAIP3). Biochem Pharmacol. 80(12):2009-20. PMID: 20599425

3.Mele A, Cervantes JR, Chien V, Friedman D, Ferran C. (2014) Single nucleotide polymorphisms at the TNFAIP3/A20 locus and susceptibility/resistance to inflammatory and autoimmune diseases. Adv Exp Med Biol. 809:163-83. PMID: 25302371

4.Das T, Chen Z, Hendriks RW, Kool M. (2018) A20/Tumor Necrosis Factor alpha-Induced Protein 3 in Immune Cells Controls Development of Autoinflammation and Autoimmunity: Lessons from Mouse Models. Front Immunol. 9:104. PMID: 29515565

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for A20/TNFAIP3 Antibody (NBP1-77533)(40)

We have publications tested in 2 confirmed species: Human, Mouse.

We have publications tested in 1 application: WB.


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WB
(6)
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Human
(4)
Mouse
(3)
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Showing Publications 1 - 10 of 40. Show All 40 Publications.
Publications using NBP1-77533 Applications Species
Feoktistova M, Makarov R, Brenji S et al. A20 Promotes Ripoptosome Formation and TNF-Induced Apoptosis via cIAPs Regulation and NIK Stabilization in Keratinocytes Cells Feb 3 2020 [PMID: 32028675] (WB, Human) WB Human
Ma J, Xiao Y, Tian B et al. Genome-wide analyses of long non-coding RNA expression profiles and functional network analysis in esophageal squamous cell carcinoma Sci Rep Jun 24 2019 [PMID: 31235759] (WB, Human) WB Human
Lim MCC, Maubach G, Sokolova O et al. Pathogen-induced ubiquitin-editing enzyme A20 bifunctionally shuts off NF-kB and caspase-8-dependent apoptotic cell death. Cell Death Differ. 2017 Jun 02 [PMID: 28574503] (WB, Human) WB Human
Reihill JA, Malcomson B, Bertelsen A et al. Induction of the inflammatory regulator A20 by gibberellic acid in airway epithelial cells. Br. J. Pharmacol. 2015 May 25 [PMID: 26013851] (WB, Human) WB Human
Xuan NT, Wang X, Nishanth G et al. A20 expression in dendritic cells protects mice from LPS-induced mortality. Eur. J. Immunol. 2014 Dec 04 [PMID: 25530110] (WB, Mouse)

Details:
A20 antibody used for WB on lysates of bone marrow-derived dendritic cells (BMDCs) isolated from CD11c-Cre A20fl/fl and A20fl/fl control mice (Figure 1A).
WB Mouse
De A, Dainichi T, Rathinam CV, Ghosh S. The deubiquitinase activity of A20 is dispensable for NF-kB signaling. EMBO Rep. 2014 May 30 [PMID: 24878851] (WB, Mouse)

Details:
Primary mouse bone marrow derived macrophages, WB: Figs 1C, 5B, 5D; IP: S9. For IP, the immunoprecipitated A20 was was used in an in vitro deubiquitinase (DUB) assay (S9). Notes: TNFa and LPS treatment upregulated A20 expression (Fig 1C). The A20 mAb reco
WB Mouse
Lai Ting-Yu, Wu Shang-Duen, Tsai Mong-Hsun et al. Transcription of Tnfaip3 is regulated by NF-kappaB and p38 via C/EBPbeta in activated macrophages. PLoS One. 2013 [PMID: 24023826] (Mouse) Mouse
Svingerud T, Solstad T, Sun B et al. Atlantic salmon type I IFN subtypes show differences in antiviral activity and cell-dependent expression: evidence for high IFNb/IFNc-producing cells in fish lymphoid tissues. J Immunol. 2012 Dec 15 [PMID: 23169587]
Karube K, Nakagawa M, Tsuzuki S et al. Identification of FOXO3 and PRDM1 as tumor-suppressor gene candidates in NK-cell neoplasms by genomic and functional analyses. Blood. 2011 Sep 22 [PMID: 21690554]
Gerlach B, Cordier SM, Schmukle AC et al. Linear ubiquitination prevents inflammation and regulates immune signalling. Nature. 2011 Mar 31 [PMID: 21455173]
Show All 40 Publications.

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Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for A20/TNFAIP3 Antibody (NBP1-77533). (Showing 1 - 3 of 3 FAQs).

  1. Following cleavage of A20/TNFAIP3, two full length fragments are released, one with 37 kDa and other one was approx. at 52 kDa. There are mentioning of 75Kda fragment and 37 KDa fragment in scientific papers but not 52 KDa. What is the significance of that specific band?
    • There is data available that explains 52 KDa band in A20/TNFAIP3. Using this antibody in Western Blot will result multiple bands. In LPS treatment, 52 KDa putative cleavage band was observed; however, the same band was not found in untreated THP1 cells.
  2. I am interested in using Mouse monoclonal A20/TNFAIP3 antibody in Mouse tissue for Immunohistochemical staining? Would I be able to achieve desired results?
    • Yes. We recommend using a directly conjugated mouse Monoclonal A20/TNFAIP3 antibody (NBP1-77533). This mouse monoclonal antibody is currently available conjugated to Phycoerythrin, Fluorescein, etc.
  3. For my experiment, I need to double stain and I would like to use A20/TNFAIP3 (59A426) Antibody NBP1-77533 for double staining with another Mouse Monoclonal Primary antibody. Is this feasible?
    • In order to double stain, we recommend a directly conjugated antibody. This antibody is available in conjugated formats, so you may be able to use either HRP, Phycoerythrin, Fluorescein or other conjugates listed in the dropdown menu.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NBP1-77533AF350
Alexa Fluor 405 NBP1-77533AF405
Alexa Fluor 488 NBP1-77533AF488
Alexa Fluor 532 NBP1-77533AF532
Alexa Fluor 594 NBP1-77533AF594
Alexa Fluor 647 NBP1-77533AF647
Alexa Fluor 700 NBP1-77533AF700
Alexa Fluor 750 NBP1-77533AF750
Allophycocyanin NBP1-77533APC
Biotin NBP1-77533B
DyLight 350 NBP1-77533UV
DyLight 405 NBP1-77533V
DyLight 488 NBP1-77533G
DyLight 550 NBP1-77533R
DyLight 594 NBP1-77533DL594
DyLight 650 NBP1-77533C
DyLight 680 NBP1-77533FR
DyLight 755 NBP1-77533IR
FITC NBP1-77533F
HRP NBP1-77533H
Janelia Fluor 549 NBP1-77533JF549
Janelia Fluor 646 NBP1-77533JF646
PE NBP1-77533PE
PerCP NBP1-77533PCP

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Bioinformatics

Gene Symbol TNFAIP3
Uniprot