HTRF Strep-Tactin®-Tb cryptate HTRF®
HTRF Strep-Tactin®-Tb cryptate can be used to capture Strep-tag® II and Twin Strep-tag®-tagged proteins.
-
No-wash
-
Ease-of-use
-
High affinity
Overview
Strep-Tactin® has been labeled with Tb crypate. 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®-Tb binds to the Strep-Tag®II tagged partner A, while partner B* binds to a specific Ab labeled with an HTRF acceptor.
*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 an acceptor for the detection.
Assay protocol
The example on the right describes the protocol using a 20 µL final assay volume for detection of 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®-Tb cryptate (5 µL) and anti-partner B labeled with acceptor (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 an acceptor for the detection.
How do the number of tests relate to the active moiety?
The average conjugate quantity per well reflects the 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 total quantity of 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.
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
Advance your CAR-T cell research - Flyers
Product Insert Strep-Tactin-Tb / 610STTLA-610STTLB
610STTLA-610STTLB - Product Insert
Batch Information Strep-Tactin-Tb / 20250831
610STTLA Batch 01A - Batch Information
Safety Data Sheet (DEU) Strep-Tactin-Tb / 610STTLA
610STTLA - Safety Data Sheet
Safety Data Sheet (ELL) Strep-Tactin-Tb / 610STTLA
610STTLA - Safety Data Sheet
Safety Data Sheet (FRA-FR) Strep-Tactin-Tb / 610STTLA
610STTLA - Safety Data Sheet
Safety Data Sheet (ITA) Strep-Tactin-Tb / 610STTLA
610STTLA - Safety Data Sheet
Safety Data Sheet (SPA) Strep-Tactin-Tb / 610STTLA
610STTLA - Safety Data Sheet
Safety Data Sheet (ENG-GB) Strep-Tactin-Tb / 610STTLA
610STTLA - Safety Data Sheet
Safety Data Sheet (ENG-US) Strep-Tactin-Tb / 610STTLA
610STTLA - Safety Data Sheet
Batch Information Strep-Tactin-Tb / 20250831
610STTLB Batch 01A - Batch Information
Safety Data Sheet (DEU) Strep-Tactin-Tb / 610STTLB
610STTLB - Safety Data Sheet
Safety Data Sheet (ELL) Strep-Tactin-Tb / 610STTLB
610STTLB - Safety Data Sheet
Safety Data Sheet (FRA-FR) Strep-Tactin-Tb / 610STTLB
610STTLB - Safety Data Sheet
Safety Data Sheet (ITA) Strep-Tactin-Tb / 610STTLB
610STTLB - Safety Data Sheet
Safety Data Sheet (SPA) Strep-Tactin-Tb / 610STTLB
610STTLB - Safety Data Sheet
Safety Data Sheet (ENG-GB) Strep-Tactin-Tb / 610STTLB
610STTLB - Safety Data Sheet
Safety Data Sheet (ENG-US) Strep-Tactin-Tb / 610STTLB
610STTLB - Safety Data Sheet
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