NF-kappaB Signaling

Phospho-CARD11 (Ser652) Antibody #5189

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Applications Key:   W = Western Blotting
Reactivity Key: H = Human  M = Mouse  Mk = Monkey  R = Rat  
Species cross-reactivity is determined by Western blot.

Protocols

5189

Western Blotting,

Specificity / Sensitivity

Phospho-CARD11 (Ser652) Antibody detects endogenous levels of CARD11 protein only when phosphorylated at Ser652. This site corresponds to Ser671 in mouse and Ser668 in rat CARD11 proteins.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to a region surrounding Ser652 of human CARD11 protein. Antibodies were purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from Jurkat cells, untreated or treated with TPA #9905 (40 nM) and calcium ionophore A23187 (2 μM) for 15 minutes, and λ-phosphatase plus calf intestinal phosphatase (CIP) as indicated, using Phospho-CARD11 (Ser652) Antibody (upper) and CARD11 (1D12) Rabbit mAb #4435 (lower).

Background

CARD11/Carma1/Bimp3 belongs to the MAGUK (membrane-associated guanylate kinase) family that typically function as molecular scaffolds in the assembly of multiprotein complexes (1,2). MAGUK family members contain an SH3 domain, a PDZ domain and a GuK domain homologous to guanylate kinase. In addition, CARD11 contains an amino-terminal CARD domain (caspase recruitment domain). This domain plays an important role in forming interactions with a number of proteins containing CARD domains that are involved in regulating apoptosis and NF-κB activation. CARD11 is predominately expressed in lymphocytes (1,2) and associates with the CARD domain of Bcl10. When overexpressed, CARD11 leads to the phosphorylation of Bcl10 and activation of NF-κB (1,2). CARD11 is constitutively associated with lipid rafts and is thought to function by recruiting Bcl10 and MALT1 and triggering the phosphorylation of IKKs (3,4). Several studies using the genetic disruption of CARD11 or dominant-negative mutations have demonstrated that it plays a critical role in NF-κB activation and lymphocyte signaling (4-7).

1. Bertin, J. et al. (2001) J. Biol. Chem.276, - .
2. Gaide, O. et al. (2001) FEBS Lett.496, 121 - 127.
3. Stilo, R. et al. (2004) J. Biol. Chem.279, 34323 - 34331.
4. Wang, D. et al. (2002) Nat. Immunol.3, 830 - 835.
5. Jun, J.E. et al. (2003) Immunity18, 751 - 762.
6. Hara, H. et al. (2003) Immunity18, 763 - 775.
7. Gaide, O. et al. (2002) Nat. Immunol.3, 836 - 843.

Application References

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Companion Products

• 4440 CARD11 Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO® Reagent and 20X Peroxide
• 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 4435 CARD11 (1D12) Rabbit mAb

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.