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HTRF Biotin-Heparan sulfate-Eu cryptate reagent, 5,000 Assay Points

A proprietary heparan sulfate substrate labeled with both biotin and Eu3+ Cryptate to monitor heparanase activity.

For research use only. Not for use in diagnostic procedures.

Product Variants
Part number: 61BHSKAA
Unit Size: 5,000 Assay Points
List price: USD 2,085.00
Your price:
USD 2,085.00
USD 2,085.00 /each
Part number: 61BHSKAB
Unit Size: 20,000 Assay Points
List price: USD 7,029.00
Your price:
USD 0.00
USD 7,029.00 /each

Overview

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.

Specifications

Assay Points
5000
Assay Target Type
Fluorescent reagent
Assay Technology
HTRF
Brand
HTRF
Quantity
1
Unit Size
5,000 Assay Points

Video gallery

References

How it works

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.

HTRF Biotin-Heparan sulfate-Eu cryptate reagent 1.svg

 

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.

HTRF Biotin-Heparan sulfate-Eu cryptate reagent 2.svg

 

Assay validation

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.

HTRF Biotin-Heparan sulfate-Eu cryptate reagent 3.svg

 

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 Biotin-Heparan sulfate-Eu cryptate reagent 4.svg
HTRF Biotin-Heparan sulfate-Eu cryptate reagent 5.svg

Resources

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Guide
HTRF solutions, guide to major applications

This guide provides you an overview of HTRF applications in several therapeutic areas.

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SDS, COAs, Manuals and more

Are you looking for technical documents for this product. We have housed them in a dedicated section., click on the links below to explore.

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