Catalog # NM5491

NLRC4 (Ser-533), phospho-specific M549 Antibody

Rat Monoclonal

Application / Dilution

ELISA	1:1000
WB	1:500

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 110 kDa

The nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) family is a diverse family of cytoplasmic innate immune receptors that are involved in recognition of pathogen-associated molecular patterns. NLRs are important for pathogen sensing, transcriptional activation of proinflammatory cytokines and activation of inflammatory caspases. NLRC4 (IPAF, CARD12) forms the inflammasome that responds to bacterial flagellin. This inflammasome is activated by NLRC4 oligomerization, NAIP protein binding, and activation of caspase-1 leading to pyroptosis. NLRC4 is phosphorylated on Ser-533 by PKCδ following infection of macrophages with S. typhimurium. Mutant forms of NLRC4 demonstrate that an unphosphorylatable form (S533A) does not activate caspase-1 and pyroptosis in response to S. typhimurium, while a phosphomimetic NLRC4 (S533D) mutant causes rapid macrophage pyroptosis without infection. Thus, PKCδ phosphorylation of NLRC4 (S533) may be a critical event in inflammasome activation and host innate immunity.

References

Qu, Y. et al. (2012) Nature 490:539.
Elinav, E. et al. (2011) Immunity 34:665.
Poyet, J.L. et al. (2001) J Biol Chem 276:28309.

Western blot image of mouse recombinant NLRC4 (Ser-533) phosphorylated peptide (lanes 1-4). The blot was dephosphorylated with lambda phosphatase (lanes 2 & 4) then the blot was probed with rat monoclonal anti-NLRC4 (Ser-533) phospho-specific (lanes 1 & 2) and rabbit polyclonal anti-NLRC4 (a.a. 525-538) (lanes 3 & 4).

Clone M549 was generated from a phospho-NLRC4 (Ser-533) synthetic peptide (coupled to carrier) corresponding to amino acids surrounding serine 533 in mouse NLRC4. This sequence has high homology to human and rat NLRC4, and is not homologous to sequences in other NLR family members.

*For more information, see UniProt Accession Q3UP24

Rat monoclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

The antibody was affinity purified using phospho-NLRC4 (Ser-533) peptide. This antibody antibody detects a 110 kDa* protein on SDS-PAGE immunoblots of PMA-differentiated THP1 human macrophages. In addition, the antibody detects a mouse recombinant phospho-NLRC4 (Ser-533) peptide, but does not detect this peptide after lambda phosphatase treatment.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

This kit contains: