PGN-ECndss ultrapure peptidoglycan

Soluble sonicated peptidoglycan from E. coli K12

PGN-ECndss is an ultrapure preparation of peptidoglycan (PGN) from the Gram-negative Escherichia coli K12.

PGN is a major component of the bacterial cell wall. It is mainly single-layered in Gram-negative bacteria (e.g. E. coli) and multi-layered in Gram-positive bacteria (e.g. B. subtilis). PGN is a large polymeric molecule made of glycan strands connected by short peptides. PGN structure varies among bacterial species at the level of sugar polymers, and of cross-linked amino-acids. In Gram-positive bacteria, the amino-acid side chain structure consists of L-alanine, D-glutamate, L-lysine, and D-alanine. In Gram-negative bacteria, the L-Lysine is replaced by meso-diaminopimelic acid (mDAP or meso-DAP) [1].

PGN, either intact or fragmented, is sensed by the innate immune system through different PRRs (pattern recognition receptors). PGN is known to be a potent activator of NF-κB and TNF-α through TLR2, although it has been reported that PGN agonist activity for TLR2 is likely relying on other commonly co-purified molecules, such as cell wall lipoproteins and lipoteichoic acids (LTAs). The role of TLR2 in the direct recognition of PGN remains controversial, and the discrepancies among studies may arise from the purification methods as well as from the variation in PGN structure in the different bacterial species [1-2].

Soluble PGN-ECndss from E. coli K12 is obtained after sonication of insoluble PGNs. At working concentrations, it activates  HEK-Blue™ NOD1 and HEK-Blue™ NOD2 cells but not HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.

KEY FEATURES OF PGN-ECndss:

• Ultrapure preparation of peptidoglycan from Escherichia coli K12
• Sonicated soluble form
• Functionally-tested using cellular reporter assays
• NOD1 and NOD2 agonist

References:

1. Wolf A.J. & Underhill D.M., 2018. Peptidoglycan recognition by the innate immune system. Nat. Rev. Immunol. 18(4):243-254.
2. Travassos.LH. et al., 2004. Toll-like receptor 2-dependent bacterial sensing does not occur via peptidoglycan recognition. EMBO Rep. 5(10):1000-1006.

Specifications

Specificity: NOD1/NOD2 agonist

Working Concentration:

• 30 - 100 µg/ml for human NOD1 in cell culture assays
• 3 - 100 µg/ml for human NOD2 in cell culture assay

Quality control:

• The biological activity of PGN-ECndss has been verified using HEK-Blue™ hNOD1 cells and HEK-Blue™ hNOD2 cells
• The absence of bacterial contaminants has been confirmed using HEK-Blue™ hTLR2 cells (for lipoproteins) and HEK-Blue™ hTLR4 cells (for endotoxins).

Contents

PGN-ECndss is provided as a solution at 1 mg/ml

• 1 ml of soluble sonicated peptidoglycan from Escherichia coli K12 (PGN-ECndss)
• 1.5 ml of sterile endotoxin-free water

PGN-ECndss is shipped at room temperature.

PGN-ECndss can be stored at room temperature (15-25°C) for 2 weeks and at 4°C for up to 6 months.

Note: When stored at 4°C, PGN-ECndss may precipitate. Place the product in a water bath at 37°C for 5 minutes and homogenize for complete solubilization.

Do not freeze. PGN-ECndss will not return to solubility.