Limulus amoebocyte lysate (LAL) is widely relied on for the detection of bacterial endotoxin. LAL is an aqueous extract of amoebocyte cells found in the Atlantic horseshoe crab from Limulus polyphemus. Gel-clot, chromogenic and turbidimetric tests are the main methods of LAL used.
LAL gel clot assay is the simplest form of LAL test and is widely used to detect the presence or absence of endotoxin in the prepared sample. [1] It initiates a series of enzymatic reactions, when endotoxin encounters LAL. The combination of endotoxin and zymogen Factor C initiates the protease cascade in the LAL test. The Factor B, which stimulated by activated Factor C, converts the proclotting enzyme to the clotting enzyme. The clotting enzyme alters the amoebocyte coagulogen that presents in LAL to form a gel-clot, which can be observed. [2] [3]
The testing should always be performed according to the manufacturer's recommendations. The endotoxin concentration is approximated by continuously applying a dilution of sample containing endotoxin to an LAL gel of different sensitivities (critical LPS amounts were known) until a negative reaction (no observable clot) is obtained. The process can take up to several hours. [4]
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