LFn-Needle - Human NLRC4 Inflammasome Inducer

T3SS Needle from B. thailandensis

ABOUT

Human NLRC4/NAIP Inflammasome Inducer - LFn-Needle

InvivoGen provides LFn-Needle, an NLRC4/NAIP inflammasome agonist. Needle is a component of the type III secretion systems (T3SS) of intracellular bacteria able to interact with NLRC4 via NAIP [1-4].

More details

 

LFn-Needle is fused to the amino-terminal domain of B. anthracis lethal factor (LFn). This fusion system, when co-administred with the anthrax toxin’s protective antigen (PA), allows intracellular delivery of the bacterial ligand [4]. The combination of LFn-Needle with the anthrax protective antigen (PA) is named Needle-Tox [5]. Its ability to activate the human NLRC4 inflammasome has been validated using THP1-NLRC4, THP1-KO-NLRC4, and THP1-Null2 cell lines. The subsequent production of IL-1β by these cells was measured using the HEK-Blue™ IL-1β cellular assay.

 

Key features:

  • Needle from B. thailandensis T3SS
  • Potent inducer of the human NLRC4 inflammasome in vitro
  • Produced in Sf9 insect cells
  • Each lot is functionally tested

 

Download our Practical guide on Inflammasomes.

 

 

References:

1. Zhao Y. et al., 2011. The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus. Nature. 477(7366):596-600.
2. Rauch I. et al., 2016. NAIP proteins are required for cytosolic detection of specific bacterial ligands in vivo. The Journal of Exp. Med. 213(5):657-665.
3. Zhao Y. et al., 2016. Genetic functions of the NAIP family of inflammasome receptors for bacterial ligands in mice. J Exp Med. 213(5):647-656.
4. Worrall L.J. et al., 2011. Structural overview of the bacterial injectisome. Curr Opin Microbiology. 14(1):3-8.
5. Ballard J.D. et al., 1996. Anthrax toxin-mediated delivery of a cytotoxic T-cell epitope in vivo. PNAS. 93(22):12531-12534.

All products are for research use only, and not for human or veterinary use.

Inflammasome activation with Needle
Inflammasome activation with Needle

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SPECIFICATIONS

Specifications

Source
Insect cells
Accession sequence

WP_009896110 (Needle sequence)

Molecular weight
~ 46 kDa (SDS-PAGE gel)
Tag
His, N-ter
Working concentration

0.16 - 100 ng/ml for cellular assays

Purity
> 90% (SDS-PAGE)
Solubility

100 μg/ml in water

Formulation buffer

150 mM
sodium chloride, 20 mM sodium phosphate buffer with 2% human
serum albumin (HSA) and 5% saccharose

Sterility

0.2 µm filtration

Tested applications

Inflammasome cellular assays

Quality control

Each lot is functionally tested and validated using THP1-Null2 cells

CONTENTS

Contents

  • Product: 
    LFn-Needle
  • Cat code: 
    tlrl-ndl
  • Quantity: 
    5 µg
Includes:

5 μg of lyophilized LFn-Needle protein
1.5 ml of endotoxin-free water

Notes:

B. anthracis protective antigen (PA) is not provided

Shipping & Storage

  • Shipping method:  Room temperature

    • Storage:

    • -20 °C

    Caution:

    • Avoid repeated freeze-thaw cycles

Details

NLRC4 BACKGROUND:

The NLRC4 (aka Ipaf) inflammasome is an intracellular multi-protein complex that plays a central role in innate immunity [1,2]. It is activated by a two-step process; a first signal (‘priming’) is provided by microbial molecules such as TLR ligands, while the second signal is provided by intracellular bacterial molecules such as Flagellin from the motility apparatus, or Inner Rod and Needle proteins from the bacterial type III or IV secretion systems (T3SS or T4SS). NLCR4 is an indirect sensor: it interacts with NAIPs (NLR family apoptosis inhibitory proteins) that directly bind to Flagellin, Needle, and Inner Rod. While a single NAIP operates upstream of NLRC4 in humans and recognizes each of these activators [3], multiple NAIPs have been described in mice with different affinities for each molecule [4-7]. The NLRC4 inflammasome appears to protect mucosal barriers, such as the lung, stomach, and intestine, from invading bacteria [2]. 

 

 

References:

1. Platnich J.M. & Muruve D.A., 2019. NOD-like receptors and inflammasomes: A review of their canonical and non-canonical signaling pathways. Arch Biochem Biophys. 670:4-14.
2. Bauer R. & Rauch I., 2020. The NAIP/NLRC4 inflammasome in infection and pathology. Mol Aspects Med. 76:100863.
3. Yang J. et al., 2013. Human NAIP and mouse NAIP1 recognize bacterial type III secretion needle protein for inflammasome activation. PNAS. 110(35):14408-14413.
4. Zhao Y. et al., 2011. The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus. Nature. 477(7366):596-600.
5. Rauch I. et al., 2016. NAIP proteins are required for cytosolic detection of specific bacterial ligands in vivo. The Journal of Exp. Med. 213(5):657-665.
6. Zhao Y. et al., 2016. Genetic functions of the NAIP family of inflammasome receptors for bacterial ligands in mice. J Exp Med. 213(5):647-656.
7. Kofoed E.M. & Vance R.E., 2011. Innate immune recognition of bacterial ligands by NAIPs determines inflammasome specificity. Nature. 477(7366):592-595.

DOCUMENTS

Documents

LFn-Needle

Technical Data Sheet

Validation Data Sheet

Safety Data Sheet

Certificate of analysis

Need a CoA ?

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