HIGH-THROUGHPUT ASSAY OF LETHAL FACTOR USING IMMOBILIZED FULL-LENGTH SUBSTRATE WITH A DEFINED ORIENTATION BASED ON IMMUNO-FLUORESCENCE METHOD

摘要

The invention of the anthrax lethal factor using immunofluorescence High-Throughput Assay relates to a method, and more Specifically, using the subcloned to cover the whole of the anthrax lethal factor and mechanism of action in the region of the substrate cleavage site and a C- terminal involved in the specific reaction between the substrate and the cleavage site can be easily observed GST (inclusive Tachikawa on -S- transmitter buffer raise) the entire structure (native form) producing a fusion protein of a substrate and a fusion protein using the glutathione (glutathione) which specifically binds the GST portion of the fusion protein to a solid support with capture ligands by fixing to allow directional, when testing the activity of the anthrax lethal factor high sensitivity (sensitivity) and specificity (specificity) as well as to the activity assay for a number of samples in a short time with a small amount of enzyme is given to the reaction After the reaction solution, and it is possible to easily remove the cutting products is improved to reduce the experimental error caused by chemicals used for screening, using the immunofluorescence of the anthrax lethal factor high-throughput relates to the activity assay method