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Lipopolysaccharide from Escherichia coli K12

LPS is the principal component of Gram-negative bacteria that activates the innate immune system. LPS recognition is predominantly mediated by Toll-like receptor 4 (TLR4), ultimately leading to the activation of NF-κB and the production of proinflammatory cytokines [1]. LPS-EK is a preparation of a rough (R)-form lipopolysaccharide (LPS) purified from the Gram-negative E. coli K12. E. coli K12 strains are the prototypical laboratory strains and are the preferred model in biochemical genetics, molecular biology, and biotechnology. LPS isolated from E. coli K12 lacks the O-antigen in its structure and is therefore considered 'rough' LPS [2].

InvivoGen provides LPS-EK with two grades of purity:

• LPS-EK is a standard lipopolysaccharide (LPS) preparation extracted by a phenol-water mixture. LPS-EK contains other bacterial components, such as lipoproteins, and therefore stimulates both TLR4 and TLR2.

• LPS-EK Ultrapure is extracted by successive enzymatic hydrolysis steps and purified by the previously described phenol-TEA-DOC extraction protocol [3]. This process removes contaminating lipoproteins, and therefore LPS-EK Ultrapure only activates TLR4. 

References:

1. Kuzmich, N.N. et al., 2017. TLR4 Signaling Pathway Modulators as Potential Therapeutics in Inflammation and Sepsis. Vaccines (Basel) 5unol. 165(2):618-22.
2. Kuhnert, P. et al., 1995. Rapid and accurate identification of Escherichia coli K-12 strains. Appl Environ Microbiol 61, 4135-4139.
3. Hirschfeld M. et al., 2000. Cutting edge: repurification of lipopolysaccharide eliminates signaling through both human and murine toll-like receptor 2. J Immunol. 165(2):618-22.

Specifications

Working concentration: 1 ng- 10 µg/ml.

LPS-EK
Specificity: TLR4 and TLR2 agonist.

LPS-EK Ultrapure
Specificity: TLR4 agonist.

Contents

• 5 mg standard lipopolysaccharide from E. coli K12 (LPS-EK)

• 1.5 ml endotoxin-free water

or

• 1 mg ultrapure lipopolysaccharide from E. coli K12 (LPS-EK Ultrapure)

• 1.5 ml endotoxin-free water

 LPS-EK is shipped at room temperature.

 LPS-EK should be stored at -20°C.
Upon resuspension, prepare aliquots of LPS-EK and store at 4°C for short term storage or -20 ̊C for long term storage.

 Lyophilized product is stable 1 year at -20°C. Resuspended product is stable 1 month at 4°C and 6 months at -20°C.

 Avoid repeated freeze-thaw cycles.

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