Flow cytometric analysis of protease activities in vital cells.

  title={Flow cytometric analysis of protease activities in vital cells.},
  author={Gregor Rothe and S Klingel and Irmgard Assfalg-Machleidt and Werner Machleidt and Christian Zirkelbach and Richard B. Banati and Walter F. Mangel and G{\"u}nter K Valet},
  journal={Biological chemistry Hoppe-Seyler},
  volume={373 7},
The analysis of lysosomal proteases in cell lysates is complicated by pH-dependent and oxidative changes of their activity and complex formation with cytosolic inhibitors. Therefore, new flow cytometric methods were developed for the intracellular measurement of protease activities in viable cells. Intracellular cleavage of substrates such as Z-Arg-Arg-4-trifluoromethylcoumarinyl-7-amide to green fluorescent 7-amino-4-trifluoromethylcoumarin (AFC) in viable neutrophils and monocytes was only… CONTINUE READING