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A20/TNFAIP3 Antibody (59A426) [Alexa Fluor® 647]

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Flow Cytometry: A20/TNFAIP3 Antibody (59A426) [Alexa Fluor® 647] [NBP1-77533AF647] - An intracellular stain was performed on SK-MEL-28 cells with A20/TNFAIP3 (59A426) antibody NBP1-77533AF647 (blue) and a matched ...read more
A20/TNFAIP3 (59A426) was detected in immersion fixed Caco-2 human colorectal adenocarcinoma cell line using Mouse anti-A20/TNFAIP3 (59A426) Protein G Purified Monoclonal Antibody conjugated to Alexa Fluor® 647 ...read more

Product Details

Summary
Reactivity Hu, Mu, RtSpecies Glossary
Applications WB, Simple Western, Flow, ICC/IF, IHC, IP, CyTOF-ready
Clone
59A426
Clonality
Monoclonal
Host
Mouse
Conjugate
Alexa Fluor 647

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A20/TNFAIP3 Antibody (59A426) [Alexa Fluor® 647] 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
  • CyTOF-ready
  • Flow (Intracellular)
  • Flow Cytometry
  • Immunocytochemistry/ Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation
  • Simple Western
  • Western Blot
Application Notes
Optimal dilution of this antibody should be experimentally determined.

Packaging, Storage & Formulations

Storage
Store at 4C in the dark.
Buffer
50mM Sodium Borate
Preservative
0.05% Sodium Azide
Purity
Protein G purified

Notes

Alexa Fluor (R) products are provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment; (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com. This conjugate is made on demand. Actual recovery may vary from the stated volume of this product. The volume will be greater than or equal to the unit size stated on the datasheet.

Alternate Names for A20/TNFAIP3 Antibody (59A426) [Alexa Fluor® 647]

  • 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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Product General Protocols

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for A20/TNFAIP3 Antibody (NBP1-77533AF647). (Showing 1 - 1 of 1 FAQ).

  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.

Secondary Antibodies

 

Isotype Controls

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Bioinformatics

Gene Symbol TNFAIP3