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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 5
#5630

A20/TNFAIP3 (D13H3) Rabbit mAb

Western Blotting Image 1: A20/TNFAIP3 (D13H3) Rabbit mAb Expand Image
Western blot analysis of extracts from G-361, JJN-3, and A20 cell lines using A20/TNFAIP3 (D13H3) Rabbit mAb.
Western Blotting Image 2: A20/TNFAIP3 (D13H3) Rabbit mAb Expand Image
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 Image 3: A20/TNFAIP3 (D13H3) Rabbit mAb Expand Image
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.
Immunoprecipitation Image 1: A20/TNFAIP3 (D13H3) Rabbit mAb Expand Image
Immunoprecipitation of A20 cell lysate using A20/TNFAIP3 (D13H3) Rabbit mAb (lane 1) or Rabbit (DA1E) mAb IgG XP® Isotype Control (lane 3). Western blot was performed using A20/TNFAIP3 (D13H3) Rabbit mAb.