Heparanase Assay
Heparanase is an enzyme that cleaves heparan sulfate thereby releasing growth factors that are implicated in tumor cell proliferation, metastasis and angiogenesis. Cisbio Bioassays's HTRF®-based assay monitors heparanase activity using a proprietary heparan sulfate substrate labeled with both biotin and Eu3+ Cryptate. This homogeneous assay has the robustness of HTRF® technology and rapidly produces high quality results.
- Homogeneous assay
- Simple and rapid assay protocol
- Amenable to high throughput automation
- 24 hour signal stability
- Miniaturization down to 10 µl
- Monitor heparanase activity
- Heparanase inhibitor screening
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 reduction in SA-XL665 emissions. The basic assay contains two phases: the heparanase enzyme reaction and HTRF detection using SA-XL665.
Product Specifications:
The effects of heparanase enzyme concentration and the enzyme reaction time on substrate degradation are shown in Figure 1. 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 for inhibitor screening.
Figure 1: 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.
Assay Performance:
An HTRF Heparanase assay standard curve was performed using the conditions outlined in the product insert (figure 2A). Assay CVs were lower than 3%. An assay using 90 ng/ml heparanase was then run in the presence of suramin, a known inhibitor of heparanases. The inhibition curve is shown in figure 2B. The calculated IC50 was 4µM which agrees with values calculated using radiometric methods (1-10µM).
Figure 2: (A) HTRF Heparanase assay was performed using a range of enzyme concentrations following standard assay conditions with a final assay volume of 20 µl. (B) The heparanase assay was performed using 90ng/ml heparanase and incubated for 30 minutes at room temperature in the presence of various concentrations of Suramin.
Ordering Info:
Heparanase Assay Reagents:
| Description | Quantity* | Cat no. |
| Biotin-Heparan-Sulfate-K | 5,000 tests | 61BHSKAA |
| Biotin-Heparan-Sulfate-K | Bulk size | on request |
| *based on 20µl assay volume | ||
Companion Products:
| Description | Quantity* | Cat no. | Product Insert |
| Streptavidin-XLent! | 5,000 tests | 611SAXLA | |
| Streptavidin- XLent! | 20,000 tests | 611SAXLB | |
| *based on 20µl assay volume | |||
For more information or To Place an Order:
U.S. and Canada
1-888-963-4567
Europe and other countries
+33(0)466 796 705
Japan
+81-3-5510-2932
