Anti-NLRC5 Antibody, clone 3H8

The protein named NLRC5, or also known as Nucleotide-binding oligomerization domain protein 27, Caterpiller protein 16.1 (CLR16.1), or Nucleotide-binding oligomerization domain protein 4 and encoded by the gene NLRC5/NOD27/NOD4, plays a role in the homeostatic control of innate immunity and anti-viral defense mechanisms. NLRC5 also likely regulates the NF kappa B and type 1 interferon signaling response pathways induced upon infection. NLRC5 interacts with CHUK and IKBKB and prevents CHUK and IKBKB phosphorylation thus inhibiting their kinase activity. NLRC5 interacts with DDX58 and IFIH1 proteins and blocks the interaction of MAVS to DDX58 proteins. NLRC5 is localized to the cytoplasm. NLRC5 is expressed in spleen, thymus, lung, brain, tonsil, heart and prostate at moderate levels, but NLRC5 expression is dramatically induced by interferons during pathological infections. Traditional immune responses studies have demonstrated that NLRC5 is a specific and master regulator of major histocompatibility complex (MHC) class I genes as well as related genes involved in MHC class I antigen presentation. The expression of MHC class I genes is regulated by NLRC5 in coordination with the RFX components through an enhanceosome-dependent manner, and the involvement of NLRC5 in MHC class I mediated CD8+ T cell activation, proliferation and cytotoxicity has proved to be critical for host defense against intracellular bacterial infections.

Target specificity of clone 3H8 was confirmed by Western blotting using lysates from cells overexpressing exogenously transfected NLRC5 as well as cells knocked down of endogenous NLRC5 by shRNA transfection (Neerincx, A., et al. (2010). J. Biol. Chem. 285(34):26223-26232). Clone 3H8 epitope is present in human NLRC5 spliced isoforms 1 and 4, but absent from isoforms 2, 3, 5, and 6 reported by UniProt (Q86WI3).

BSA-conjugated linear peptide corresponding to the C-terminal sequence of human NLRC5.

Detect NLRC5 using this rat monoclonal antibody, Anti-NLRC5 Antibody, clone 3H8 validated for use in western blotting, IP & Flow Cytometry.

Flow Cytometry Analysis: 0.25 µg from a representative lot detected NLRC5 in Jurkat and Raji cells.

Immunoprecipitation Analysis: A representative lot detected poly(I:C) (Cat. No. 528906) treatment-induced upregulation of endogenous NLRC5 in HeLa cells by IP-Western blotting analysis of both cytosolic and nuclear fractions. A strong NLRC5 nuclear accumulation was seen upon nuclear export inhibition by LepB (Cat. No. 431050) treatment, whereas NLRC5 was mainly cytoplasmic in untreated cells (Neerincx, A., et al. (2012). J. Immunol. 188(10):4940-4950).

Western Blotting Analysis: A representative lot detected NLRC5 in THP-1 cell lysate as well as exogenously expressed FLAG-tagged NLRC5 in lysate from transfected HEK293T cells. Target band detection was greatly diminished using lysates from NLRC5 siRNA-transfected THP-1 cells. (Neerincx, A., et al. (2010). J. Biol. Chem. 285(34):26223-26232).

Research Category
Inflammation & Immunology

Research Sub Category
Immunoglobulins & Immunology

Evaluated by Western Blotting in Raji cell lysate.

Western Blotting Analysis: 1 µg/mL of this antibody detected NLRC5 in 10 µg of Raji cell lysate.

~205 kDa observed. 204.6 kDa (isoform 1) and 201.3 kDa (isoform 4) calculated. Uncharacterized band(s) may appear in some lysates.

Protein G Purified

Purified rat monoclonal IgG1κ in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Stable for 1 year at 2-8°C from date of receipt.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.