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Significance Researchers and clinicians rely on robust staining methods to detect and visualize biomolecules of interest. Mucins are heavily glycosylated proteins expressed on cells throughout the human body that have historically proved challenging to stain with high specificity, hindering attempts to explore their roles in health and disease. Here, we address this limitation using catalytically inactivated mucin-cleaving bacterial proteases as staining reagents. The differing specificities of these enzymes are retained in their binding properties, enabling a new depth in the analysis of mucins on cells and patient tissues.

An enzymatic toolkit for selective proteolysis, detection, and visualization of mucin-domain glycoproteins