Biotin-Heparan sulfate-Eu cryptate reagent HTRF®

A proprietary heparan sulfate substrate labeled with both biotin and Eu3+ Cryptate to monitor heparanase activity.
See more
  • No-wash No-wash
  • Ease-of-use Ease-of-use
A proprietary heparan sulfate substrate labeled with both biotin and Eu3+ Cryptate to monitor heparanase activity.
In stock


HTRF heparan sulfate substrate labeled with both biotin and Eu3+ Cryptate used with HTRF SA-XL665, allows to monitor heparanase activity with the robustness of HTRF technology.

Heparanase is an enzyme that cleaves heparan sulfate, and thereby releases growth factors that are implicated in tumor cell proliferation, metastasis and angiogenesis.



Assay principle

The HTRF heparanase assay is based on our proprietary heparan sulfate substrate, labeled with both biotin and Eu3+ Cryptate. Active heparanase enzyme cleaves the substrate causing the loss of energy transfer, and thus a reduction in SA-XL665 emissions.
assay principle of heparanase assay

Assay protocol

The assay is run as a two-step process: 1.Heparanase enzymatic reaction: Mix together, the substrate, the heparanase and/or compounds, incubate for 30 min at 37°C (We recommend a time course study to determine the optimal stimulation time) 2.Heparanase activity detection: Add Streptavidin-XL665
assay protocol of heparanase assay

Heparanase assay: enzymatic kinetic

HTRF Heparanase assay was performed using a range of enzyme concentrations. The heparanase enzyme reaction was allowed to proceed for various times prior to adding SA-XL665 and the emissions detected. The effects of heparanase enzyme concentration and the enzyme reaction time on substrate degradation are shown here. Based on the linearity of the enzyme dose-response, enzyme consumption and incubation time results, 90 ng/mL of heparanase incubated for 30 minutes at 37ºC was chosen for inhibitor screening.
Heparanase assay using HTRF labeled-Heparan substrate and SaXL665

Heparanase assay: inhibitory effect of suramin

Heparanase assay was performed using a range of enzyme concentrations with a final assay volume of 20 µL. Then an inhition assay was performed using heparanase at 90ng/mL incubated for 30 minutes at RT in presence of various concentrations of Suramin.

Fig A shows the Heparanase assay standard curve, assay CVs were lower than 3%. Fig.B shows the inhibitory effect of Suramin, a known inhibitor of heparanases. The IC50 was calculated at 4 µM, which is in agreement with values calculated using radiometric methods (1-10µM).

HTRF heparanase assay titration curve
Inhibitory effect of suramin on HTRF heparanase assay

HTRF PPI your dream assay served on a plate

Sandwiches aren't just for eating - Infographics

Best practices for pharmacological characterization of PPI inhibitors

Easy pharmacological characterization of PPI modulators. - Technical Notes

HTRF assays handle low- to high affinity protein-protein interactions

Deciphering low- and high affinity interactions - Application Notes

Nuclear receptor ligand identification with HTRF

Monitoring nuclear receptor binding with HTRF assays - Application Notes

HTRF addresses large protein-protein interaction complexes

Challenge large complexes with HTRF assays - Application Notes

A brief history of Protein-Protein Interactions

How well do you know PPI? - Infographics

HTRF Product Catalog

All your HTRF assays in one document! - Catalog

A guide to Homogeneous Time Resolved Fluorescence

General principles of HTRF - Guides

How HTRF compares to Western Blot and ELISA

Get the brochure about technology comparison. - Brochures

Virology research solutions using HTRF Protein-Protein Interaction assays

See how peer researchers challenge the viral life cycle with PPI assays - Application Notes

Plate Reader Requirement

Choosing the right microplate reader ensures you’ll get an optimal readout. Discover our high performance reader, or verify if your lab equipment is going to be compatible with this detection technology.

Let's find your reader

Latest news