Description
Overview
Strep-Tactin has been labeled with d2. Biotin, Twin Strep-tag,and Strep-tagII 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.
HTRF assays offer many advantages over other technologies:
Homogeneous add-and-read format
No wash steps
Low background
Straightforward miniaturization from 96- or 384-well microplates to high density assay formats such as 384-well low volume and 1536-well plates
Stable signal, providing flexibility in time of readout or size of assays
How it works
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®-d2 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 a donor for the detection.
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®-d2 (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.
