Our NLRC4 (Ser-533) rabbit polyclonal phosphospecific primary antibody from PhosphoSolutions is produced in-house. It detects human, mouse, and rat NLRC4 (Ser-533) and is antigen affinity purified. It is great for use in WB.
Western blot image of mouse recombinant NLRC4 (Ser-533) phosphorylated peptide (lanes 1 & 4) and dephosphorylated peptide (lanes 2 & 5), as well as human PMA-differentiated THP1 cells (lanes 3 & 6). The blots were probed with rabbit polyclonals anti-NLRC4 (a.a. 525-538) (lanes 1-3) and anti-NLRC4 (Ser-533) phospho-specific (lanes 4-6).
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.
Antigen Affinity Purified
Polyclonal
IgG
WB
Rabbit
NLRC4
110
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.
Human, Mouse, Rat
Storage at -20°C is recommended, as aliquots may be taken without freeze/thawing due to presence of 50% glycerol. Stable for at least 1 year at -20°C.
Liquid
PBS + 1 mg/ml BSA, 0.05% NaN3 and 50% glycerol
WB: 1:1000
Unconjugated
The antibody was cross-adsorbed to unphosphorylated NLRC4 (Ser-533) peptide before affinity purification 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.
Phosphorylated
Ser-533
Western blots performed on each lot.
For research use only. Not intended for therapeutic or diagnostic use. Use of all products is subject to our terms and conditions, which can be viewed on our website.
United States
After date of receipt, stable for at least 1 year at -20°C.
Blue Ice
CARD12, CLAN1, IPAF, NLR family CARD domain-containing protein 4, NOD-like receptor 4
Mohamed, MF, et al. 2022. CrkII/Abl phosphorylation cascade is critical for NLRC4 inflammasome activity and is blocked by Pseudomonas aeruginosa ExoT. Nature communications, 1295.
Paudel, S, et al. 2019. NLRC4 suppresses IL-17A-mediated neutrophil-dependent host defense through upregulation of IL-18 and induction of necroptosis during Gram-positive pneumonia. Mucosal immunology, 247-257.
Liu, W, et al. 2017. LRRK2 promotes the activation of NLRC4 inflammasome during Salmonella Typhimurium infection. The Journal of experimental medicine, 3051-3066.
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