Revision 5

#5630Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

82

Source/Isotype:

Rabbit IgG

UniProt ID:

#P21580

Entrez-Gene Id:

7128

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

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

Species Reactivity:

Human, Mouse, Rat, Monkey

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.

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.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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