Related Experiment Video
Updated: Nov 15, 2025
16:02
Quantitative FRET Förster Resonance Energy Transfer Analysis for SENP1 Protease Kinetics Determination
Published on: February 21, 2013
19.4K
A Robust, One-step FRET Assay for Human Heparanase
Jyothi C Sistla1,2, Umesh R Desai1,2
1Institute for Structural Biology, Drug Discovery, and Development, Virginia Commonwealth University, Richmond, VA 23219, USA.
Bio-Protocol
|March 3, 2021
Summary
A novel heparanase assay using FRET technology offers a simpler, high-throughput method for screening inhibitors and measuring enzyme activity in biological samples.
Area of Science:
- Biochemistry
- Enzymology
- Molecular Biology
Background:
- Heparanase (endo-β-D-glucuronidase) cleaves heparan sulfate (HS) chains, influencing cell growth and metastasis.
- Existing heparanase assays are often complex, labor-intensive, and costly.
Purpose of the Study:
- To develop a simplified, high-throughput heparanase assay.
- To utilize Förster Resonance Energy Transfer (FRET) with Dabcyl and EDANS labeled heparin.
Main Methods:
- Heparin was labeled with donor (Dabcyl) and acceptor (EDANS) fluorophores.
- The FRET signal generated upon heparin cleavage by heparanase was measured.
- Assay robustness and applicability for inhibitor screening were evaluated.
Main Results:
- A FRET-based assay for heparanase activity was successfully developed.
- The assay demonstrates potential for robust and high-throughput screening of heparanase inhibitors.
- The assay can be applied to study heparanase activity in tumors and biological fluids.
Conclusions:
- A simplified FRET-based assay provides a robust method for heparanase activity measurement.
- This assay facilitates high-throughput screening of potential heparanase inhibitors.
- The assay is suitable for analyzing heparanase in various biological contexts, including oncology and diagnostics.
