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Cord Factor (endotoxin-free grade)

Immunomodulator
 
ALX-581-210-M001 1 mg 215.00 USD
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Immunomodulatory compound / adjuvant. Induces activation of macrophages independent of TLR2 or TLR4.

Product Specification

Alternative Name:Trehalose 6,6’-dimycolate, TDM, 6,6’-Di-O-mycoloyl-α,α-trehalose
 
Source:Isolated from M. tuberculosis.
 
CAS:61512-20-7
 
Formulation:Lyophilized.
 
Purity:Single spot (TLC)
 
Endotoxin Content:<0.0002EU/µg (LAL test; BioWhittaker)
 
Solubility:Soluble in chloroform:methanol:water (90:10:1) (5mg/ml), hexane or isopropanol.
 
Shipping:Ambient
 
Long Term Storage:+4°C
 
Protocol:In vitro stimulation of macrophages
Purified Cord Factor was used to stimulate either mouse RAW 264.7 cells or bone marrow-derived macrophages. Cord Factor was suspended at a concentration of 1mg/ml in isopropanol and sonicated in a bath sonicator for 5 min. This suspension was incubated at 60°C for 10 min. and sonication repeated. The resulting solution was layered onto 24-well tissue culture plates at the indicated concentrations and incubated at 37°C in order to ensure complete evaporation of the solvent. Control wells were layered with solvent without Cord Factor and incubated at 37°C. To this layer of Cord Factor, either RAW 264.7 cells or bone marrow-derived macrophages were added at a concentration of 106 cells in 100µl of medium and incubated at 37°C for 24 hours before activation e.g. TNF-α production was measured in the supernatant.

Alternatively, Cord Factor was suspended at a concentration of 0.2 or 2mg/ml in hexane. Of the resulting solution 50µl were layered onto 96-well tissue culture plates at the indicated concentrations of 1 or 10µg/well, respectively and the solvent completely evaporated. Control wells were layered with solvent without Cord Factor and incubated at 37°C. To this layer of Cord Factor, either RAW 264.7 cells or bone marrow-derived macrophages were added at a concentration of 106 cells in 100µl of medium and incubated at 37°C for 24 hours before activation e.g. TNF-α production was measured in the supernatant.

In vivo pulmonary granuloma formation in mice
For in vivo experiments 10µg Cord Factor per mouse were applied i.v. in a water/oil/water emulsion.
 
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Product Literature References

MARCO variants are associated with phagocytosis, pulmonary tuberculosis susceptibility and Beijing lineage: N.T. Thuong, et al.; Genes Immun. 17, 419 (2016), Abstract;
Heat shock protein 90 inhibition abrogates TLR4-mediated NF-κB activity and reduces renal ischemia-reperfusion injury: S. O'Neill; Sci. Rep. 5, 12958 (2015), Application(s): In vitro experiment; Reagent , Abstract;
Interferon-gamma independent formation of pulmonary granuloma in mice by injections with trehalose dimycolate (cord factor), lipoarabinomannan and phosphatidylinositol mannosides isolated from Mycobacterium tuberculosis: H. Takimoto, et al.; Clin. Exp. Immunol. 144, 134 (2006), Abstract;
Macrophage scavenger receptor down-regulates mycobacterial cord factor-induced proinflammatory cytokine production by alveolar and hepatic macrophages: Y. Ozeki, et al.; Microb. Pathog. 40, 171 (2006), Abstract;
In vivo activity of released cell wall lipids of Mycobacterium bovis bacillus Calmette-Guerin is due principally to trehalose mycolates: R.E. Geisel, et al.; J. Immunol. 174, 5007 (2005), Abstract;
Requisite role for complement C5 and the C5a receptor in granulomatous response to mycobacterial glycolipid trehalose 6,6’-dimycolate: C.W. Borders, et al.; Scand. J. Immunol. 62, 123 (2005), Abstract;
Failure of CD1D-/- mice to elicit hypersensitive granulomas to mycobacterial cord factor trehalose 6,6’-dimycolate: T.V. Guidry, et al.; J. Interferon Cytokine Res. 24, 362 (2004), Abstract;
Cord factor trehalose 6,6’-dimycolate (TDM) mediates trafficking events during mycobacterial infection of murine macrophages: J. Indrigo, et al.; Microbiology 149, 2049 (2003), Abstract;
Influence of trehalose 6,6’-dimycolate (TDM) during mycobacterial infection of bone marrow macrophages: J. Indrigo, et al.; Microbiology 148, 1991 (2002), Abstract;
Mycobacterial glycolipid cord factor trehalose 6,6’-dimycolate causes a decrease in serum cortisol during the granulomatous response: J.K. Actor, et al.; Neuroimmunomodulation 10, 270 (2002), Abstract;
Immunological properties of trehalose dimycolate (cord factor) and other mycolic acid-containing glycolipids: R. Ryll, et al.; Microbiol. Immunol. 45, 801 (2001), Review, Abstract;
In vivo administration of mycobacterial cord factor (Trehalose 6, 6'-dimycolate) can induce lung and liver granulomas and thymic atrophy in rabbits: N. Hamasaki, et al.; Infect. Immun. 68, 3704 (2000), Abstract;
Extravascular coagulation and fibrinolysis in murine lung inflammation induced by the mycobacterial cord factor trehalose-6,6’-dimycolate: R.L. Perez, et al.; Am. J. Respir. Crit. Care Med. 149, 510 (1994), Abstract;
Development of a trehalose 6,6’-dimycolate model which explains cord formation by Mycobacterium tuberculosis: C.A. Behling, et al.; Infect. Immun. 61, 2296 (1993), Abstract;
Induction of resistance to tuberculosis in mice with defined components of mycobacteria and with some unrelated materials: E. Ribi, et al.; Immunology 46, 297 (1982), Abstract;
The chemical structure of the cord factor of Mycobacterium tuberculosis: H. Noll, et al.; Biochim. Biophys. Acta 20, 299 (1956), Abstract;

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