Detection of protease activity by FRET using porous silicon as an energy acceptor

✍ Scribed by Gu, Luo ;Orosco, Manuel ;Sailor, Michael J.

• Book ID: 105365237
• Publisher: John Wiley and Sons
• Year: 2009
• Tongue: English
• Weight: 239 KB
• Volume: 206
• Category: Article
• ISSN: 0031-8965
• DOI: 10.1002/pssa.200881065

✦ Synopsis

Abstract

This work demonstrates a method for detection of protease activity using porous Si as the energy acceptor in a Fluorescence Resonance Energy Transfer (FRET) assay. A fluorescent dye (fluorescein) is trapped in a porous Si matrix, where its fluorescence is quenched. The dye is trapped using a protein (zein), and the action of proteases (Pronase E) release the dye from the quenching matrix. A strong fluorescence signal appears within 30 min of Pronase E addition. A control with no Pronase E shows little fluorescence, and a control using heat‐denatured Pronase E shows approximately 50% of the fluorescence of the active protease. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)