-->

HTRF Strep-Tactin®-XL665 HTRF®

HTRF Strep-Tactin®-XL665 can be used to capture Strep-tag® II and Twin Strep-tag®-tagged proteins.

See more
  • No-wash No-wash
  • Ease-of-use Ease-of-use
  • High affinity High affinity

HTRF Strep-Tactin®-XL665 can be used to capture Strep-tag® II and Twin Strep-tag®-tagged proteins.

-

Overview

Strep-Tactin® has been labeled with XL665. Biotin, Twin Strep-tag®,and Strep-tag®II bind to Strep-Tactin® with high affinity. The binding is rapid and stable, making it an ideal choice for use in a variety of assays such as enzyme assays, protein-protein binding assays, and molecular biology assays.

Benefits

  • LARGE COMPLEX DETECTION
  • BATCH-TO-BATCH REPRODUCIBILITY
  • LOW TO HIGH KD DETECTION

Assay principle

In an HTRF interaction assay, one partner is labeled (directly or indirectly) with the donor, and the other with the acceptor (again, directly or indirectly). The intensity of the signal is proportional to the binding of the 2 partners. In the example shown here, Strep-Tactin®-XL665 binds to the Strep-Tag®II tagged partner A ,while partner B* binds to a specific Ab labeled with an HTRF donor.

*partner B can also be biotinylated, tagged, Fc fused. In these cases, use the corresponding HTRF reagent (anti-Tag, anti-species, protA, Streptavidin) labeled with donor for the detection.

Principle of the HTRF protein-protein interaction assay Strep-tactin XL665

Assay protocol

The example on the right describes the protocol using a 20 µL final assay volume for detecting an interaction between a Strep-Tag®II-tagged partner A and a non-tagged partner B*. Dispense the 2 partners (10 µL), incubate, add Strep-Tactin®-XL665 (5 µL) and anti-partner B labeled with donor (5 µL), incubate, and then read.

*partner B can also be tagged, Fc fused, or directly labeled. In these cases use the corresponding HTRF reagent (anti-Tag, anti-species, protA, Streptavidin), labeled with a donor for the detection.

Protocol of the HTRF protein-protein interaction assay Strep-tactin XL665

Build HTRF interaction assays for your specific application

Reagents are sold by the number of tests (20 µL reactions). XL665 or d2 conjugates are supplied on the basis of 40 ng of Strep-Tactin® per well. The amount of active moiety per vial is also indicated (as well as the number of tracers per vial - see product description sheet). The active moiety is defined as the active part of a conjugate (e.g. Strep-Tactin®).

How do the number of tests relate to the active moiety?

The average conjugate quantity per well reflects overall biological material content. Using the active moiety amount is generally preferred to the quantity of total conjugate. For Cryptate and d2 conjugates, the total conjugate amount equals that of the active moiety, since the molecular weight of the label is negligible. This is not the case for XL665 labeled entities, for which the quantity of total conjugate will vary depending on the final molar ratio of the XL665 conjugate. However, the amount of active moiety indicated by Cisbio is constant and based on the number of tests ordered.

Active Moiety

Recommended quantities of Cryptate and XL665 conjugates

Cryptate conjugates must not be excessive, in order to prevent reader saturation and an unacceptable level of background. In most cases, a cryptate concentration of 1 to 5nM is appropriate, and will generate 20,000 to 80,000 cps at 620 nm depending on the HTRF compatible reader used. The XL665 conjugate must match its assay counterpart as closely as possible in order for the maximum number of biomolecules to be tagged with the XL665 acceptor. Thus, to detect a tagged molecule at an assay concentration of 20nM, the concentration of anti-Tag-XL665 should be equimolar or higher.

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

HTRF PPI your dream assay served on a plate

Sandwiches aren't just for eating - Infographics

A brief history of Protein-Protein Interactions

How well do you know PPI? - Infographics

Virology research solutions using HTRF Protein-Protein Interaction assays

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

Solutions for CAR-T research

Advance your CAR-T cell research - Flyers

Batch Information Strep-Tactin-XL665 / 20250907

610STXLA Batch 01A - Batch Information

Batch Information Strep-Tactin-XL665 / 20250907

610STXLB Batch 01A - Batch Information

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