Revision 1

#12421Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

110

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q9NPP4

Entrez-Gene Id:

58484

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

NLRC4 (D5Y8E) Rabbit mAb recognizes endogenous levels of total NLRC4 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu942 of human NLRC4 protein.

Background

The nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) family of proteins is a diverse family of cytoplasmic innate immune receptors. They are characterized by the presence of an amino-terminal effector domain, which is often either a caspase activation and recruitment domain (CARD) or a pyrin domain (PYD), followed by a NACHT domain and carboxy-terminal leucine-rich-repeats (LRR) involved in recognition of pathogen-associated molecular patterns (PAMPs) (1). NLR proteins play a variety of roles during the innate immune response including pathogen sensing, transcriptional activation of proinflammatory cytokines through NF-κB, transcriptional activation of type I interferons through IRFs, and formation of inflammasomes leading to activation of inflammatory caspases (1-7).
The NLRC4 (IPAF) inflammasome forms in response to bacterial flagellin as well as components of the bacterial conserved type II secretion system (TTSS) (8-12). Ligand detection and ligand-dependent NLRC4 oligomerization and inflammasome activation require the NAIP family of proteins (13,14). In mice, NAIP5 and NAIP6 associate with flagellin, while NAIP2 interacts with TTSS rod proteins (13,14). In humans, NAIP recognizes the TTSS needle protein Cprl (14). In addition, NLRC4 is phosphorylated by PKCδ in response to bacterial infection and this phosphorylation is required for inflammasome assembly and caspase-1 activation (15).

  1. Elinav, E. et al. (2011) Immunity 34, 665-79.
  2. Inohara, N. et al. (1999) J Biol Chem 274, 14560-7.
  3. Ogura, Y. et al. (2001) J Biol Chem 276, 4812-8.
  4. Sabbah, A. et al. (2009) Nat Immunol 10, 1073-80.
  5. Mariathasan, S. et al. (2004) Nature 430, 213-8.
  6. Agostini, L. et al. (2004) Immunity 20, 319-25.
  7. Martinon, F. et al. (2002) Mol Cell 10, 417-26.
  8. Poyet, J.L. et al. (2001) J Biol Chem 276, 28309-13.
  9. Mariathasan, S. et al. (2004) Nature 430, 213-8.
  10. Franchi, L. et al. (2006) Nat Immunol 7, 576-82.
  11. Miao, E.A. et al. (2006) Nat Immunol 7, 569-75.
  12. Miao, E.A. et al. (2010) Proc Natl Acad Sci U S A 107, 3076-80.
  13. Kofoed, E.M. and Vance, R.E. (2011) Nature 477, 592-5.
  14. Zhao, Y. et al. (2011) Nature 477, 596-600.
  15. Qu, Y. et al. (2012) Nature 490, 539-42.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

Revision 1
#12421

NLRC4 (D5Y8E) Rabbit mAb

Western Blotting Image 1: NLRC4 (D5Y8E) Rabbit mAb Expand Image
Western blot analysis of extracts from THP-1 and MUTZ-3 cells using NLRC4 (D5Y8E) Rabbit mAb.
Western Blotting Image 2: NLRC4 (D5Y8E) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human NLRC4 (hNLRC4; +), using NLRC4 (D5Y8E) Rabbit mAb.
Immunoprecipitation Image 1: NLRC4 (D5Y8E) Rabbit mAb Expand Image
Immunoprecipitation of NLRC4 from MUTZ-3 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or NLRC4 (D5Y8E) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using NLRC4 (D5Y8E) Rabbit mAb.