Accedi

Arthur G. James Cancer Hospital and Richard J. Solove Research Institute

2 ARTICLES PUBLISHED IN JoVE

Cancer Research

Detection of Protease Activity by Fluorescent Peptide Zymography

Ameya A. Deshmukh^1,2, Jessica L. Weist¹, Jennifer L. Leight^1,2

¹Comprehensive Cancer Center, James Cancer Hospital and Solove Research Center, Ohio State University, ²Department of Biomedical Engineering, College of Engineering, Ohio State University

Here, we present a detailed protocol for a modified zymographic technique in which fluorescent peptides are used as the degradable substrate in place of native proteins. Electrophoresis of biological samples in fluorescent peptide zymograms enables detection of a wider range of proteases than previous zymographic techniques.

Bioengineering

Measuring Global Cellular Matrix Metalloproteinase and Metabolic Activity in 3D Hydrogels

Abdulaziz S. Fakhouri^1,2,3, Jennifer L. Leight^1,2

¹Department of Biomedical Engineering, The Ohio State University, ²The Ohio State University Comprehensive Cancer Center, Arthur G. James Cancer Hospital and Richard J. Solove Research Institute, ³Biomedical Technology Department, King Saud University

Here, a protocol is presented for encapsulating and culturing cells in poly(ethylene glycol) (PEG) hydrogels functionalized with a fluorogenic matrix metalloproteinase (MMP)-degradable peptide. Cellular MMP and metabolic activity are measured directly from the hydrogel cultures using a standard microplate reader.