Author: Larouche-Lebel, Éva; Loughran, Kerry A.; Oyama, Mark A.; Solter, Phil F.; Laughlin, Danielle S.; Sánchez, Melissa D.; Assenmacher, Charles-Antoine; Fox, Philip R.; Fries, Ryan C.
Title: Plasma and tissue angiotensin-converting enzyme 2 activity and plasma equilibrium concentrations of angiotensin peptides in dogs with heart disease Document date: 2019_6_28
ID: 6ek80l79_1
Snippet: The assay utilizes a synthetic peptide conjugate as a reaction substrate that is cleaved by ACE2 to release a free fluorophore. The fluorescence (Ex/Em=320 nm/420 nm) of each sample is quantified every 8 min for up to 120 min by a 96-well microplate fluorescence reader. Change in relative fluorescence units over time is converted to the amount of 4-methylcoumaryl-7-amide (MCA) released per minute using a standard curve generated by measuring the .....
Document: The assay utilizes a synthetic peptide conjugate as a reaction substrate that is cleaved by ACE2 to release a free fluorophore. The fluorescence (Ex/Em=320 nm/420 nm) of each sample is quantified every 8 min for up to 120 min by a 96-well microplate fluorescence reader. Change in relative fluorescence units over time is converted to the amount of 4-methylcoumaryl-7-amide (MCA) released per minute using a standard curve generated by measuring the endpoint fluorescence (Ex/Em =320/420 nm) of known concentrations of a MCA standard solution ranging from 0 to 250 pmol/well of MCA. One mU of ACE2 activity is defined as the release of 1 pmol of MCA from the substrate per min.
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