UNIT 13.12 Microsphere-Based Flow Cytometry Protease Assays for Use in Protease Activity Detection and High-Throughput Screening

  1. Matthew J. Saunders1,2,3,
  2. Bruce S. Edwards2,
  3. Jingshu Zhu1,
  4. Larry A. Sklar2,
  5. Steven W. Graves1,3

Published Online: 1 OCT 2010

DOI: 10.1002/0471142956.cy1312s54

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Saunders, M. J., Edwards, B. S., Zhu, J., Sklar, L. A. and Graves, S. W. 2010. Microsphere-Based Flow Cytometry Protease Assays for Use in Protease Activity Detection and High-Throughput Screening. Current Protocols in Cytometry. 54:13.12:13.12.1–13.12.17.

Author Information

  1. 1

    Center for Biomedical Engineering, Department of Chemical and Nuclear Engineering, University of New Mexico, Albuquerque, New Mexico

  2. 2

    University of New Mexico Center for Molecular Discovery, Albuquerque, New Mexico

  3. 3

    National Flow Cytometry Resource, Los Alamos National Laboratory, Los Alamos, New Mexico

Publication History

  1. Published Online: 1 OCT 2010
  2. Published Print: OCT 2010


This protocol describes microsphere-based protease assays for use in flow cytometry and high-throughput screening. This platform measures a loss of fluorescence from the surface of a microsphere due to the cleavage of an attached fluorescent protease substrate by a suitable protease enzyme. The assay format can be adapted to any site or protein-specific protease of interest and results can be measured in both real time and as endpoint fluorescence assays on a flow cytometer. Endpoint assays are easily adapted to microplate format for flow cytometry high-throughput analysis and inhibitor screening. Curr. Protoc. Cytom. 54:13.12.1-13.12.17. © 2010 by John Wiley & Sons, Inc.


  • protease assays;
  • microspheres;
  • high-throughput;
  • protease;
  • multiplex microspheres;
  • green fluorescent protein