Source

Lipopolysaccharide (LPS) from E. coli strain EH100 (Ra), R-type (rough/mutant) LPS

Concentration

1mg/ml stabilised in sterile, double-distilled water (ddWater), without any additives

TLRpure^TM

No detectable TLR4 independent activity as determined by a mouse macrophage cell culture cytokine secretion assay using TLR4 deficient versus wild-type cells: standardised potent TLR4-specific agonist

Purity

Ultrapure. No detectable DNA, RNA and protein traces.

Purification Method

R-type (mutant/rough) LPS was isolated by phenol-chloroform-petroleum-ether method. Semi-purified LPS was subjected to further re-extraction cycles and ultracentrifugation steps, extensively electrodialysed to yield TLRpure^TM LPS.

Sterility

Filter method: certified according to Ph. Eur. 9.

Passed according to specification:

• No growth in Thioglycolate medium at 30-35°C after 14 days.

• No growth in Soybean Casein Digest Broth (TSB) at 20-25°C after 14 days.

Endotoxin Content

Bacterial Endotoxin Test (kinetic turbidimetric LAL method) certified according to Ph. Eur. 9.

Endotoxin Content: >10,000,000 [EU/ml].

Appearance

Colourless, clear, aqueous solution

Handling

Keep sterile. Prepare working dilutions from pre-warmed (~40°C) LPS stock solution just prior to use. Ready-made solution is cell culture-grade. To yield a 100µg/ml (1,000-100x) stock solution add 100µl of LPS to 900µl endotoxin-free and sterile ddWater (Cat. No.: IAX-900-002) or 0.9% NaCl Solution (Cat. No.: IAX-900-003) and mix well.

Activity

Optimal concentration is dependent upon cell type, species, desired activation and analysis:
0.01-1.0µg/ml. Does not activate any TLR other than TLR4 as tested up to 1µg/ml in relevant cellular systems (mouse macrophages).