HTRF® - Homogenous Time-Resolved Fluorescence

HTRF® is a TR-FRET based technology from Cisbio Bioassays that uses the principles of both TRF and FRET. The HTRF® donor fluorophore is either Europium cryptate (Eu3+ cryptate) or Lumi4™-Tb (Tb2+ cryptate), fruit of a recent collaboration with Lumiphore Inc.. Both donors have the long-lived emissions of lanthanides coupled with the stability of cryptate encapsulation. XL665, a modified allophycocyanin, is the HTRF® primary acceptor fluorophore. d2 represents a second generation of acceptor characterized by an organic structure 100 times smaller.

 

HTRF® is a highly flexible chemistry and has been successfully used to measure molecular complexes of many different sizes. This includes assessment of small phosphorylated peptides, immunoassays for quantifying large glycoproteins such as thyroglobulin, receptor tyrosine kinase activity using membrane preparations and indirect detection (via secondary antibodies) of tagged complexes such as CD28/CD86 binding (see figure to the left).

 

 

BMG LABTECH has published numerous Application Notes (AN) that demonstrate the use of HTRF® technology with microplate readers including the PHERAstar Series and the Omega Series.

HTS Instrument Discovers Low Affinity Inhibitors of the Inositol Phosphate (IP) Signaling Pathway(AN 222)
Functional Screening of recombinant CHO-M1 cells using the PHERAstar from BMG LABTECH (AN 193)
HTRF®IP-One Terbium-based assay performed on BMG LABTECH´s PHERAstar Plus (AN 177)
Cisbio´s HTRF® cortisol assay performed on the PHERAstar (AN 154)
HTRF® IP-One Assay Performed on the PHERAstar and RUBYstar Plate Readers (AN 137)
A novel PI 3-Kinase assay utilising HTRF® technology on the RUBYstar (AN127)
PHERAstar: A Next Generation Multimode Plate Reader for Sophisticated HTS Assays (AN 126)

The following BMG LABTECH microplate readers are certified HTRF® compatible from Cisbio Bioassays:

For more information about HTRF® and the TR-FRET technology, please see this website: www.htrf.com

The fluorescence ratio associated with HTRF® readout is a correction method developed by CIS bio international and covered by the US patent 5,527,684 and its foreign equivalents. The dual simultaneous fluorescence time-resolved measurement is covered by CIS bio international’s US patent 6,352,672 and its foreign equivalents. CIS bio international has granted BMG LABTECH a license for the promotion of these methods with its microplate readers. The application of said methods is strictly limited to the use of HTRF® reagents and technology, excluding any and all other TR-FRET technologies.