Lipopolysaccharide from Escherichia coli 055:B5

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) and ultimately leads to the activation of NF-κB and the production of proinflammatory cytokines [1]. LPS-B5 is a preparation of smooth (S)-form LPS from the Gram-negative bacteria E. coli 055:B5. LPS-B5 is the prototypical endotoxin and is often used as an endotoxin standard in Limulus amebocyte lysate (LAL) assays. In particular, LPS-B5 is highly pyrogenic and a potent activator of TLR4 [2].

InvivoGen provides LPS-B5 with two grades of purity:

• LPS-B5 is a standard lipopolysaccharide (LPS) preparation extracted by a phenol-water mixture. LPS-B5 contains other bacterial components, such as lipoproteins, and therefore stimulates both TLR4 and TLR2.

• LPS-B5 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-B5 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. Dogan M. et al., 2000. Effects of different serotypes of Escherichia coli lipopolysaccharides on body temperature in rats. Life Sci. 67(19):2319-29.
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

Specificity:
LPS-B5 (Standard): TLR2/TLR4 agonist
LPS-B5 Ultrapure: TLR4 agonist

Working concentrations :
LPS-B5 (Standard):
• TLR4 activity: 100 pg - 1 μg/ml
• TLR2 activity: 100 ng/ml - 1mg/ml
LPS-B5 Ultrapure:
• TLR4 activity: 100 pg - 1 μg/ml

Quality control :
- The TLR4 activity is validated using HEK-Blue™ TLR4 cells.
- The presence of other bacterial components (e.g. lipoproteins) is monitored using HEK-Blue™ TLR2 cells.

Contents

LPS-B5 (Standard):
- 5 mg LPS-B5 (lipopolysaccharide from E. coli serotype 055:K59(B5)H-)
Source strain: ATCC 12014; CDC 5624-50 [NCTC 9701]
- 1.5 ml endotoxin-free water

LPS-B5 Ultrapure:
- 5 mg LPS-B5 Ultrapure (Lipopolysaccharide from E. coli serotype 055:K59(B5)H-)
Source strain: ATCC 12014; CDC 5624-50 [NCTC 9701]
- 1.5 ml endotoxin-free water

 LPS-B5 is shipped at room temperature

 Stored at -20 °C.

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