Cell Signaling Technology

Product Pathways - NF-kB Signaling

A20/TNFAIP3 (D13H3) Rabbit mAb #5630

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M R Mk Endogenous 82 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

A20/TNFAIP3 (D13H3) Rabbit mAb recognizes endogenous levels of total A20/TNFAIP3 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus terminus of human A20/TNFAIP3 protein. The antibody was prepared in collaboration with Dr. Marc Schmidt-Supprian, Max Planck Institute of Biochemistry.

Western Blotting

Western Blotting

Western blot analysis of extracts from G-361, JJN-3, and A20 cell lines using A20/TNFAIP3 (D13H3) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or treated with TPA #4174 (40 nM) and A23187 (2 μM) for various amounts of time, using A20/TNFAIP3 (D13H3) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS-7 cells, mock transfected (-) or transfected with a Myc-DDK-tagged human A20/TNFAIP3 construct (+), using A20/TNFAIP3 (D13H3) Rabbit mAb.


Background

A20, also referred to as TNF-α-induced protein 3 (TNFAIP3), is cytokine-inducible protein that functions to inhibit apoptosis and activate NF-κB (1,2). It was first identified as a TNF-α inducible primary response gene in human umbilical vein endothelial cells, and encodes a 790-amino acid protein containing seven Cys2/Cys2-zinc finger motifs (3). Constitutive expression of A20 is observed in lymphoid tissues (4), but it is transiently expressed in a variety of cell types in response to inflammatory signals such as TNF-α (3,5), IL-1 (3,5), phorbol esters (6), and LPS (7). Expression of A20 can confer resistance to apoptosis and NF-κB activation triggered by these signals, probably through interference with TRAF (TNF receptor associated factor) family members (8,9), and interaction with the NF-κB inhibiting protein ABIN (10). Studies also show that A20 contains site-specific ubiquitin modifying activity that can contribute to its biological functions (11,12). The amino-terminus of A20 contains de-ubiquitinating (DUB) activity for Lys63 branches, such as those found in TRAF6 and RIP, while the carboxyl-terminus contains ubiquitin ligase (E3) activity for Lys48 branches of the same substrates and leads to their degradation (12).

  1. Beyaert, R. et al. (2000) Biochem Pharmacol 60, 1143-51.
  2. Lee, E.G. et al. (2000) Science 289, 2350-4.
  3. Dixit, V.M. et al. (1990) J Biol Chem 265, 2973-8.
  4. Tewari, M. et al. (1995) J Immunol 154, 1699-706.
  5. Jäättelä, M. et al. (1996) J Immunol 156, 1166-73.
  6. Laherty, C.D. et al. (1993) J Biol Chem 268, 5032-9.
  7. Hu, X. et al. (1998) Blood 92, 2759-65.
  8. Song, H.Y. et al. (1996) Proc Natl Acad Sci USA 93, 6721-5.
  9. Heyninck, K. and Beyaert, R. (1999) FEBS Lett 442, 147-50.
  10. Heyninck, K. et al. (1999) J Cell Biol 145, 1471-82.
  11. Evans, P.C. et al. (2004) Biochem J 378, 727-34.
  12. Lin, S.C. et al. (2008) J Mol Biol 376, 526-40.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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