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X-GlcA, 5-Bromo-4-chloro-3-indolyl-ß-D-glucuronic acid is a substrate for ß-D-Glucuronidase (GUS) encoded by the gusA gene.

The substrate is used as a qualitative histochemical marker of specific GUS expression in cells and tissue. X-GlcA is cleaved by GUS at the B1 glucuronic bond between glucuronic acid and the 5-Bromo-4-chloro-3-indolyl part of X-GlcA via hydrolysis. The cleavage of X-GlcA results in the precipitation of water insoluble blue dichloro-dibromo-indigo precipitate at the site of enzymatic cleavage.

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