Mast Cell Degranulation Assay Kit (Sigma–Aldrich) were used to quantify levels of tryptase47 (link). The kit detects the chromophore p-nitroaniline (pNA) cleaved from the labeled substrate tosyl-gly-pro-lys-pNA. In brief, the lacrimal gland lysates and the ocular surface tear wash were incubated with 0.1 mg/mL tosyl-gly-pro-lys-pNA (substrate) for 2 hours at 37 °C. A SpectraMax Plus 384 Microplate Reader (Molecular Devices, San Jose, CA, USA) was used to quantify free pNA at 405 nm.
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