Phospho-NLRC4 (Ser533) Antibody - #AF3580

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Product: Phospho-NLRC4 (Ser533) Antibody
Catalog: AF3580
Description: Rabbit polyclonal antibody to Phospho-NLRC4 (Ser533)
Application: WB IF/ICC
Cited expt.: WB
Reactivity: Human, Mouse
Mol.Wt.: 117kD(Calculated).
Uniprot: Q3UP24
RRID: AB_2846894

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MSDS
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WB Handbook
IHC Protocol
IF/ICC Protocol

Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user. For optimal experimental results, antibody reuse is not recommended.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse
Clonality:
Polyclonal
Specificity:
Phospho-NLRC4 (Ser533) Antibody detects endogenous levels of NLRC4 only when phosphorylated at Ser533.
RRID:
AB_2846894
Cite Format: Affinity Biosciences Cat# AF3580, RRID:AB_2846894.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Immunogens

Immunogen:

A synthesized peptide derived from mouse NLRC4 around the phosphorylation site of Ser533.

Uniprot:

Q3UP24(NLRC4_MOUSE) >>Visit HPA database.

Expression:
Q3UP24 NLRC4_MOUSE:

Expressed by intestinal mononuclear phagocytes.

Sequence:
MNFIRNNRRALIQRMGLTVTKQICDDLFALNVLNNQEANVIYCEPLEQEAARKIIHMTMQKGSAACNLFLKSLENWDYFVYQDLTGQNLSYQVTEEDLNVLAQNLKDLYNSPAFLNFYPLGEDIDIIFNLEKTFTEPIMWKKDHRHHRVEQLTLGSLLEALKSPCLIEGESGKGKSTLLQRIAMLWASGGCRALKGFRLVFFIHLRSARGGLFETLYDQLLNIPDFISKPTFKALLLKLHKEVLFLLDGYNEFHPQNCPEIEALIKENHRFKNMVIVTTTTECLRHIRHVGALTAEVGDMTEDSAKDLIEAVLVPDQVERLWAQIQESRCLRNLMKTPLFVVITCAIQMGRQEFQAHTQTMLFQTFYDLLIQKNSHRYRGGASGDFARSLDYCGDLALEGVFAHKFDFEPEHGSSMNEDVLVTIGLLCKYTAQRLKPTYKFFHKSFQEYTAGRRLSSLLTSKEPEEVSKGNSYLNKMVSISDITSLYGNLLLYTCGSSTEATRAVMRHLAMVYQHGSLQGLSVTKRPLWRQESIQSLRNTTEQDVLKAINVNSFVECGINLFSESMSKSDLSQEFEAFFQGKSLYINSENIPDYLFDFFEYLPNCASALDFVKLDFYERATESQDKAEENVPGVHTEGPSETYIPPRAVSLFFNWKQEFKTLEVTLRDINKLNKQDIKYLGKIFSSATNLRLHIKRCAAMAGRLSSVLRTCKNMHTLMVEASPLTTDDEQYITSVTGLQNLSIHRLHTQQLPGGLIDSLGNLKNLERLILDDIRMNEEDAKNLAEGLRSLKKMRLLHLTHLSDIGEGMDYIVKSLSEESCDLQEMKLVACCLTANSVKVLAQNLHNLIKLSILDISENYLEKDGNEALQELIGRLGVLGELTTLMLPWCWDVHTSLPKLLKQLEGTPGLAKLGLKNWRLRDEEIKSLGEFLEMNPLRDLQQLDLAGHCVSSDGWLYFMNVFENLKQLVFFDFSTEEFLPDAALVRKLSQVLSKLTLLQEVKLTGWEFDDYDISAIKGTFKLVTA

Research Backgrounds

Function:

Key component of inflammasomes that indirectly senses specific proteins from pathogenic bacteria and fungi and responds by assembling an inflammasome complex that promotes caspase-1 activation, cytokine production and macrophage pyroptosis. The NLRC4 inflammasome is activated as part of the innate immune response to a range of intracellular bacteria. It senses pathogenic proteins of the type III secretion system (T3SS) and type IV secretion system (T4SS) such as flagellin and PrgJ-like rod proteins via the Naip proteins (Naip1, Naip2 or Naip5): specific Naip proteins recognize and bind pathogenic proteins, driving assembly and activation of the NLRC4 inflammasome. The NLRC4 inflammasome senses Gram-negative bacteria such as L.pneumophila and P.aeruginosa, enteric pathogens S.typhimurium (Salmonella) and S.flexneri and fungal pathogen C.albicans. In intestine, the NLRC4 inflammasome is able to discriminate between commensal and pathogenic bacteria and specifically drives production of interleukin-1 beta (IL1B) in response to infection by Salmonella or P.aeruginosa. In case of L.pneumophila infection the inflammasome acts by activating caspase-7.

PTMs:

Phosphorylated at Ser-533 following infection of macrophages with S.typhimurium (Salmonella). Phosphorylation is essential for NLRC4 inflammasome function to promote caspase-1 activation and pyroptosis. PRKCD phosphorylates Ser-533 in vitro.

Subcellular Location:

Cytoplasm>Cytosol. Inflammasome.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Expressed by intestinal mononuclear phagocytes.

Family&Domains:

In an autoinhibited form the C-terminal leucine-rich repeat (LRR) domain is positioned to sterically occlude one side of the NBD domain and consequently sequester NLRC4 in a monomeric state. An ADP-mediated interaction between the NBD and the WHD also contributes to the autoinhibition (PubMed:23765277).

References

1). p16INK4a Deletion Alleviated Obesity-Associated Kidney Fibrosis by Regulating Metabolic Reprogramming and the Inflammasome Pathway. Journal of cellular and molecular medicine, 2025 (PubMed: 40079088) [IF=5.3]

Application: WB    Species: Mouse    Sample: renal tissues

FIGURE 3 p16 knockout (p16−/−) ameliorates the NLRP3 inflammasome pathway. (A–E) Expression levels and statistical figures of ASC, Caspase-1, NLRC4 and NLRP3 in renal tissues from 20 months ApoE−/− mice and DKO mice induced by HFD were detected by western blotting (n = 4); (F–H) Fluorescence quantitative PCR experiments were performed to detect the transcription levels of ASC, NLRC4 and NLRP3 in the renal tissues (n = 12); (I, K) expression levels and statistical analysis of ASC and NLRP3 in renal tissues from 20 months ApoE−/− mice and DKO mice detected by immunohistochemical staining (n = 3); values are mean ± SEM, p 
2). Sesamin-mediated high expression of BECN2 ameliorates cartilage endplate degeneration by reducing autophagy and inflammation. Aging, 2024 (PubMed: 38284902) [IF=3.9]

Application: WB    Species: Rat    Sample:

Figure 3. Increased BECN2 attenuated the autophagy and inflammation of LPS-induced ATDC5 degeneration. (A) Western blot was employed to detect the expression of ATG14, VPS34 and GASP1. (B) The protein expression of ATG14, VPS34 and GASP1 were determined using ImageJ software, GAPDH was used as the internal control, respectively (n=3). (C) ATG14, VPS34 and GASP1 expression were determined by immunofluorescence staining (scale bar: 100μm). (D–F) Average optical density was calculated by ImageJ software. (G) Western blot was employed to detect the expression of NLRP3, NLRC4, NLRP1 and AIM2. (H) The protein expression of NLRP3, NLRC4, NLRP1 and AIM2 were determined using ImageJ software, GAPDH was used as the internal control, respectively (n=3). (I) NLRP3, NLRC4, NLRP1 and AIM2 expression were determined by immunofluorescence staining (scale bar: 100μm). (J–M) Average optical density was calculated by ImageJ software. All data represent mean ± SD. All in vitro experiments were repeated three times independently. * p < 0.05, ** p < 0.01, and *** p < 0.001.

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