A20/TNFAIP3 Antibody (59A426) - BSA Free

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

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.

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

We have publications tested in 2 applications: ICC/IF, WB.

Showing Publications 1 - 10 of 42. Show All 42 Publications. Collapse Publications.

Publications using NBP1-77533	Applications	Species
Wang yT, Liu Ty, Shen CH et al. K48/K63-linked polyubiquitination of ATG9A by TRAF6 E3 ligase regulates oxidative stress-induced autophagy Cell reports 2022-02-22 [PMID: 35196483] (WB, ICC/IF, Mouse, Human)	WB, ICC/IF	Mouse, Human
Tran VT, Ju H Fluorescence Based on Surface Plasmon Coupled Emission for Ultrahigh Sensitivity Immunoassay of Cardiac Troponin I Biomedicines 2021-04-21 [PMID: 33919217]
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 2020-02-03 [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 2019-06-24 [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. 2017-06-02 [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. 2015-05-25 [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. 2014-12-04 [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. 2014-05-30 [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. 2013-01-01 [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. 2012-12-15 [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. 2011-09-22 [PMID: 21690554] Details: WB: Fig 2A (NKL cells over expressing A20 in an Tet-Off inducible system). Note, the specificity of the A20 antibody, clone 59A426 was validated using A20 expressed in an Tet-Off indicible system by WB (Fig 2A).
Gerlach B, Cordier SM, Schmukle AC et al. Linear ubiquitination prevents inflammation and regulates immune signalling. Nature. 2011-03-31 [PMID: 21455173] Details: WB: Fig 1b (HeLa). Note, the A20 antibody recognized A20 co-IP'd with a TNF antibody as well as A20 from total HeLa cell lysate.
Hovelmeyer N, Reissig S, Xuan NT et al. A20 deficiency in B cells enhances B-cell proliferation and results in the development of autoantibodies. Eur J Immunol. 2011-03-01 [PMID: 21341261] Details: WB (Fig 1C), purified B cells from control and A20 knockout mice. Note: The specificity of the A20 antibody was validated in A20 knockout mice by WB in Fig 1C.
Adrianto I, Wen F, Templeton A et al. Association of a functional variant downstream of TNFAIP3 with systemic lupus erythematosus. Nat Genet. 2011-03-01 [PMID: 21336280] Details: WB (Fig 3 G, H), Cell lines from donors of European or American American ancestry.
Imbeault M, Ouellet M, Giguere K et al. Acquisition of host-derived CD40L by HIV-1 in vivo and its functional consequences in the B-cell compartment. J Virol. 2011-03-01 [PMID: 21177803] Details: A20/TNFAIP3 (IMG-161A). WB: Tonsillar B cells, Fig 3B.
Malinverni C, Unterreiner A, Staal J et al. Cleavage by MALT1 induces cytosolic release of A20. Biochem Biophys Res Commun. 2010-10-01 [PMID: 20804738] Details: A20/TNFAIP3 (IMG-161A). WB: HEK293 cell line, Fig 1B.
Frenzel LP, Claus R, Plume N et al. Sustained NF-kappaB activity in chronic lymphocytic leukemia is independent of genetic and epigenetic alterations in the TNFAIP3 (A20) locus. Int J Cancer. 2011-05-15 [PMID: 20669229] Details: WB: B cells isolated from healthy blood donors and from patients with CLL, Fig 3.
Harada Y, Kanehira M, Fujisawa Y et al. Cell-permeable peptide DEPDC1-ZNF224 interferes with transcriptional repression and oncogenicity in bladder cancer cells. Cancer Res. 2010-07-15 [PMID: 20587513] Details: WB (Fig 5A): UM-UC-3 human bladder cancer cell line.
Hjelmeland AB, Wu Q, Wickman S et al. Targeting A20 Decreases Glioma Stem Cell Survival and Tumor Growth. PLoS One 2010-01-01 [PMID: 20186265]
Hailfinger S, Lenz G, Ngo V et al. Essential role of MALT1 protease activity in activated B cell-like diffuse large B-cell lymphoma. Proc Natl Acad Sci U S A. 2009-11-24 [PMID: 19897720] Details: Fig 1B, Diffuse large B-cell lymphoma (DLBCL) of the activated B-cell (ABC) subtype cell lines. DLBCL-ABC is the least curable subtype of DLBCL. A20 (~72 and higher kDa forms) and A20 cleavage fragments (~65, 58, and 47 kDa) were detected. The pattern and intensity of A20 cleavage fragments varied between cell lines. A20 cleavage was partially inhibited by a MALT1 inhibitor. 2. Fig 1C, CARMA transfected BJAB cells. BJAB is a germinal center B cell-like (GCB) DLBCL. 3. Fig S3, Jurkat, DLBCL-ABC, and GCB-ABC cell lines. A20 and A20 cleavage fragments were assessed as in Fig 1B. 37 kDa and 65 kDa A20 cleavage fragments were induced in PMA + ionomycin treated Jurkat cells.
Metellus P, Coulibaly B, Nanni I et al. Prognostic impact of O6-methylguanine-DNA methyltransferase silencing in patients with recurrent glioblastoma multiforme who undergo surgery and carmustine wafer implantation: a prospective patient cohort. Cancer. 2009-10-15 [PMID: 19637364] Details: IHC(P): human glioblastoma multiforme, results described but not shown.
Honma K, Tsuzuki S, Nakagawa M et al. TNFAIP3/A20 functions as a novel tumor suppressor gene in several subtypes of non-Hodgkin lymphomas. Blood. 2009-09-17 [PMID: 19608751] Details: WB: Normal human peripheral blood B cells (CB33) treated wtih A20 and Lucifarase siRNA, Fig 5A. Note: mAb A20 (IMG-161A) was siRNA validated by western blot, Fig 5A.
Duwel M, Welteke V, Oeckinghaus A et al. A20 negatively regulates T cell receptor signaling to NF-kappaB by cleaving Malt1 ubiquitin chains. J Immunol. 2009-06-15 [PMID: 19494296] Details: WB: Jurkat & Jurkat transfected with control or A20-specific siRNA (Figs 1B-F, 2, 4, 6A, 7A-B, 9A-C); human T cells isolated from peripheral blood and expanded (Fig 9A). IP/WB: Jurkat (Fig 6B) Notes: 1. The specificity of the A20 was A20 SiRNA validated by WB in Jurkat cells (multiple figures, see publication for details). 2. An A20 cleavage fragment (A20p37) was detected at ~37 kDa in stimulated Jurkat cells by IP/WB (Fig 6) and WB (Fig 7).
Schmitz R, Hansmann ML, Bohle V et al. TNFAIP3 (A20) is a tumor suppressor gene in Hodgkin lymphoma and primary mediastinal B cell lymphoma. J Exp Med. 2009-05-11 [PMID: 19380639] Details: Fig 1, Hodgkin lymphoma (HL) and primary mediastinal B cell lymphoma (PMBL) cell lines. A20 expression was detected in the L-428 (highest expression), DEV, and L-591 HL lines, but not in several other HL cells lines nor the Karpas PMBL cell line. Loss of A20 protein expression was found to be correlated with A20 gene mutations. 2. Fig S3, HL cell line that expressed A20 (L-428) and HL cell line that lacked detectable A20 expression (L-1236). Introduction of WT A20 into the L-1236 cell line resulted in A20 protein expression. Note: The specificity of the A20 antibody was demonstrated in Fig 1 by WB in lymphoma cells by showing that the antibody detected A20 in cells with WT but not mutant A20 genes. Additionally, the specificity of the A20 antibody was transfected validated in Fig S3 by WB where introduction of WT A20 into an A20 mutant cell line resulted in detection of A20 protein expression.
Verhelst K, Verstrepen L, Coornaert B et al. Cellular expression of A20 and ABIN-3 in response to Toll-like receptor-4 stimulation. Methods Mol Biol. 2009-01-01 [PMID: 19378032] Details: WB (Fig 1): LPS-induced A20 protein expression in THP-1 cells. Cells were left untreated or stimulated for 3, 6, 9, 12, or 24 hr with 100 ng/ml LPS.
Breckpot K, Aerts-Toegaert C, Heirman C et al. Attenuated expression of A20 markedly increases the efficacy of double-stranded RNA-activated dendritic cells as an anti-cancer vaccine. J Immunol. 2009-01-15 [PMID: 19124729] Details: WB in human DC (PBMC isolated from healthy donors were used as a source of DC precursors): 1. Fig 1b, immature DCs matured with poly(I:C) for 0-48 hr 2. Fig 1d, immature DC (lane 1), mature DC (lane 2), immature DC treated with A20 siRNA (lane 3) or irrelevant siRNA (lane 4). Note: The A20 antibody was siRNA validated in Fig 1d by WB.
Li L, Soetandyo N, Wang Q et al. The zinc finger protein A20 targets TRAF2 to the lysosomes for degradation. Biochim Biophys Acta. 2009-02-01 [PMID: 18952128] Details: A20 (IMG-161A): WB (HeLa cells), Fig 7C. Note: The expression of A20 was upregulated in HeLa cells when recombinant TWEAK was added to the cell culture media. A20 was slightly upregulated when chloroquine(lysosome inhibitor) was added to the cell culture media.
Werner SL, Kearns JD, Zadorozhnaya V et al. Encoding NF-kappaB temporal control in response to TNF: distinct roles for the negative regulators IkappaBalpha and A20. Genes Dev. 2008-08-01 [PMID: 18676814] Details: WB: 1. Fig S1B & S1C (HeLa cells treated with TNF). TNF (1 ng/ml) dramatically upregulated A20 expression in HeLa cells. Refer to the publication (page S7) for a signal pathway cartoon illustrating the TNF induction of A20 through the NF-kB signaling pathway 2. Fig S2E [MEF cells retrovirally transduced with contitutively or inducibly (through TNF treatment) constructs encoding human A20)] Note: The specificity of the A20 antibody was A20 retrovirally (human A20 construct) validated in MEF by WB in Fig S2E.
Li L, Hailey DW, Soetandyo N et al. Localization of A20 to a lysosome-associated compartment and its role in NFkappaB signaling. Biochim Biophys Acta. 2008-06-01 [PMID: 18329387] Details: A20 (IMG-161) WB, 293T cells transfected with wild type and various A20 variant constructs, Fig 6B. Note: The A20 antibody was transfected validated by western blot in 293T cells expressing recombinant wild type and variant A20 protein.
Coornaert B, Baens M, Heyninck K et al. T cell antigen receptor stimulation induces MALT1 paracaspase-mediated cleavage of the NF-kappaB inhibitor A20. Nat Immunol. 2008-03-01 [PMID: 18223652] Details: Cell lines include: Jurkat, Raji, SSK41. Primary human T cells, HEK293T and Jurkat cells transiently or stably transfected with A20 or other plasmids. Treatments include cycloheximide, PMA + isonomycin, anti-CD3 + anti-CD28 antibody and others. See Figs for details. Notes: 1. A 37 kDa A20 cleavage fragment was observed following T cell and B cell activation (Fig 1, S1 and other Figs). 2. The specificity of the A20 antibody was validated by western blot in HEK293T transfected with human A20 C-terminal or N-terminal constructs (Fig S2). The results showed that the A20 antibody (clone 59A524) recognizes an epitope in the C-terminal part of A20 (amino acids 440-790).
Chevillard G, Derjuga A, Devost D et al. Identification of interleukin-1beta regulated genes in uterine smooth muscle cells. Reproduction. 2007-12-01 [PMID: 18042638] Details: WB: Human uterine smooth (myometrial) cell line stimulated with IL-1beta, Fig 4A.
Bohgaki M, Tsukiyama T, Nakajima A et al. Involvement of Ymer in suppression of NF-kappaB activation by regulated interaction with lysine-63-linked polyubiquitin chain. Biochim Biophys Acta. 2008-05-01 [PMID: 18029035] Details: IF (Fig 1): HeLa cells; IP (Fig 3B, top blot): combinations of A20, UbK63, & Ymer recombinant proteins. The in vitro binding assay that that A20 associates with the K63 polyubiquitin chain and that association is enhanced in the presence of Ymer; IP/WB (Fig 3B, bottom blot) recombinant proteins as described for Fig 3B, top blot; WB: Figs 1B (Jurkat) and 6B (HeLa). Note: The specificity of the A20 antibody was validated in Fig 3B by WB and IP/WB using baculovirus expressed recombinant A20 protein.
Zhang B, Guan CC, Chen WT et al. A20 inhibits human salivary adenoid cystic carcinoma cells invasion via blocking nuclear factor-kappaB activation. Chin Med J (Engl). 2007-10-20 [PMID: 18028781] Details: WB [AAC-2 (human salivary adenoid cystic carcinoma cell line), ACC-2 cells stably transfected with A20 or control vector)], Fig 2. ACC-2 cells had a basal level of A20 expression which was increased in the ACC-2-A20 cells, and overexpression of A20 in the ACC-2-A20 cells corresponded to a reduction in NF-kB activity, cell growth and invasion. Note: The specificity of the A20 antibody was A20 transfected validated by WB in ACC-2 cells in Fig 2.
Yokota S, Okabayashi T, Yokosawa N et al. Measles virus P protein suppresses Toll-like receptor signal through up-regulation of ubiquitin-modifying enzyme A20. FASEB J. 2008-01-01 [PMID: 17720800] Details: WB (Fig 2): human THP-1, U937, OSC70 cell lines. A20 was upregulated in the monocytic cell lines (U937 and THP-1) but not in the epithelial cell lines (SiHa and OSC700) during measles virus infection.
Hutti JE, Turk BE, Asara JM et al. IkappaB kinase beta phosphorylates the K63 deubiquitinase A20 to cause feedback inhibition of the NF-kappaB pathway. Mol Cell Biol. 2007-11-01 [PMID: 17709380] Details: Fig 3 (WB): A20-Flag transfected HEK293T cells (Fig 3B) cells and A20-Flag retrovirally stably transduced A20-/- mouse embryo fibroblasts (MEFs) (Fig 3D). A20-Flag was immunoprecipitated with an anti-Flag antibody and immunoprecipitates were western blotted with the A20 aMab. Fig 4A (IP/WB): THP-1 cells (differentiated with PMA and then treated with LPS) were immunoprecipitated with the A20 antibody and immunoprecipitates were western blotted with the A20 mAb. Fig 5A (WB): A20-Flag transfected HEK293T cells. Fig 6C (IP/WB): A20-Flag MEFs as described for Fig 3. Note: The specificity of the A20 antibody was A20 transfected retrovirally transduced validated by in HEK293T (Figs 3B, 5A) and MEF (Fgis 3D, 5A, 6C) cells by WB.
Salaun B, Lebecque S, Matikainen S et al. Toll-like receptor 3 expressed by melanoma cells as a target for therapy? Clin Cancer Res. 2007-08-01 [PMID: 17671143] Details: WB (Fig 1): Me 260 and Me 300 human melanoma cell lines treated with IFNa, Poly(I:C) or IFNa + Poly(I;C). Cells had a low or undetectable basal level of A20. A20 expression was upregulated in Me 260 with treatment, most notably with IFNa followed by Poly(I:C).
Wang YY, Li L, Han KJ et al. A20 is a potent inhibitor of TLR3- and Sendai virus-induced activation of NF-kappaB and ISRE and IFN-beta promoter. FEBS Lett. 2004-10-08 [PMID: 15474016] Details: WB, U937 cells treated with TNF or infected with Sendai Virus, Fig 5.
Ke B, Shen XD, Lassman CR et al. Cytoprotective and antiapoptotic effects of IL-13 in hepatic cold ischemia/reperfusion injury are heme oxygenase-1 dependent. Am J Transplant. 2003-09-01 [PMID: 12919086] Details: WB: Samples were obtained 3, 7, and 14 days after rats underwent orthotopic liver transplantation (OLT), Fig 4A.
Yang KY, Arcaroli J, Kupfner J et al. Involvement of phosphatidylinositol 3-kinase gamma in neutrophil apoptosis. Cell Signal. 2003-02-01 [PMID: 12464394] Details: WB: Lung and bone marrow neutrophils from C57/BL6 (PI3Ky +/+ and PI3Ky -/-) mice, Fig 1A.
Ke B, Shen XD, Zhai Y et al. Heme oxygenase 1 mediates the immunomodulatory and antiapoptotic effects of interleukin 13 gene therapy in vivo and in vitro. Hum Gene Ther. 2002-10-10 [PMID: 12396617] Details: Fig 5A, rat cardiac allograft tissue. Fig 7, HUVEC (human umbilical cord vein) cells. Note: In Figs 5A and 7, A20 expression was upregulated through various treatments, see publication for details.
Wu WS, Xu ZX, Chang KS. The promyelocytic leukemia protein represses A20-mediated transcription. J Biol Chem. 2002-08-30 [PMID: 12080044] Details: A20 (IMG-161A). WB: Osteosarcoma stable cell lines pMEP4/U20S and PML/U2OS treated with Cd2+ and TNFalpha, Fig 1B.
Kupfner JG, Arcaroli JJ, Yum HK et al. Role of NF-kappaB in endotoxemia-induced alterations of lung neutrophil apoptosis. J Immunol. 2001-12-15 [PMID: 11739525] Details: WB, mouse lung neutrophils, Fig 2. WB, mouse peripheral neutrophils pretreated with BMS 205820 or left untreated, Fig 5.
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Array NBP1-77533

• A20/TNFAIP3 Antibodies
• A20/TNFAIP3 Lysate
• A20/TNFAIP3 Proteins