Magellan nanoquant infinite m200 pro

The assays were performed in 96-well plates in a total reaction volume of 100 µL using 50 mM Tris-HCl pH 8.0, 150 mM NaCl, 0.01% Tween-20, 0.01% BSA for dilution of all samples. 5 nM purified active recombinant HATL5 or matriptase serine protease domain was incubated at 37°C for 60 min with 100 µm of the synthetic peptide L-1720 Suc-Ala-Ala-Pro-Arg-pNA (Bachem, Bubendorf, Switzerland) in the absence or presence (inhibitor and substrate added concomitantly) of HAI-1 (60 nM) (R&D, Minneapolis, MN), HAI-2 (40 nM) (R&D, Minneapolis, MN), aprotinin (2 µm), leupeptin (20 µm), benzamidine (2 mM), or serpinA1 (60 nM) (all from Thermo Scientific, Waltham, MA). Changes in absorbance at 405 nm were monitored using a Magellan NanoQuant Infinite M200 Pro plate reader (Tecan US, Inc., Morrisville, NC).