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Keywords:

  • 78.55.Mb;
  • 87.14.ej;
  • 87.64.kv;
  • 87.85.fk

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)