Microsphere-based flow cytometry protease assays for use in protease activity detection and high-throughput screening.

Microsphere-based flow cytometry protease assays for use in protease activity detection and high-throughput screening. Curr Protoc Cytom. 2010 Oct;Chapter 13:Unit 13.12.1-17 Authors: Saunders MJ, Edwards BS, Zhu J, Sklar LA, Graves SW Abstract 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. PMID: 20938917 [PubMed - indexed for MEDLINE]
Source: Current Protocols in Cytometry - Category: Molecular Biology Tags: Curr Protoc Cytom Source Type: research