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Mild acid hydrolysis to induce selective cleavage of KDO-lipid A linkage yielded a -labeled product that partitioned during Bligh/Dyer extraction and migrated during thin-layer chromatography like lipid A.
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Chemical hydrolysis yielded -labeled free fatty acids characteristic of Francisella lipid A but with a different molar ratio of 3-OH C18:0 to 3-OH C16:0 and different composition of non-hydroxylated fatty acids (mainly C14:0 rather than C16:0) than that of free Francisella lipid A. Autoradiographic and immunologic analysis confirmed that this purified material was largely devoid of low molecular weight LPS and of the copious amounts of free lipid A that the Francisellae accumulate.
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Capsule was purified by two independent methods and yielded similar results. To pursue the possibility that the capsule of Francisella live vaccine strain (LVS) has a structurally unique lipid anchor, we have metabolically labeled Francisella with acetate to facilitate highly sensitive compositional analysis of capsule-associated lipids. Francisella tularensis, the Gram-negative bacterium that causes tularemia, produces a high molecular weight capsule that is immunologically distinct from Francisella lipopolysaccharide but contains the same O-antigen tetrasaccharide.