Homogenous time-resolved fluorescence (HTRF)

HTRF ® is a TR-FRET based technology from Cisbio. This technology uses the principles of both TRF and FRET. The HTRF ® donor fluorophore is either Europium cryptate (Eu 3+ cryptate) or Lumi4 ™ -Tb (Tb 2+ cryptate). (Lumi4 ™ -Tb (Tb 2+ cryptate) results from collaboration between Cisbio and 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 that measures biomolecular complexes of many different sizes. This includes assessment of small phosphorylated peptides, immunoassays for quantifying large glycoproteins, 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 right).

HTRF products

The following BMG LABTECH microplate readers are certified HTRF ® -compatible by Cisbio:

BMG LABTECH microplate reader PHERAstar FSX from the front view

PHERAstar FSX

Powerful and most sensitive HTS plate reader
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BMG LABTECH microplate reader CLARIOstar Plus from the front view

CLARIOstar Plus

Most flexible Plate Reader for Assay Development
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Frontview of the FLUOstar Omega, a microplate reader from the BMG LABTECH Omega series

Omega series

Upgradeable single and multi-mode microplate reader series
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HTRF measurements

BMG LABTECH has published numerous Application notes using HTRF ® technology. This technology works well on many of our readers, including the CLARIOstar and PHERAstar FSX and Omega series .

HTRF IP-One terbium cellular assay principle

Cellular and biochemical HTRF assays measured in 1536-well microplates

One assay technology, one device, and less than one second to detect 1536 reactions
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Shematic description of the epigeneous methyltransferase assay kit

Miniaturization of an HTRF methyltransferase assay that detects histone modifying activity

Read how post-translational modifiers are studied in 2 µl reaction volume
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Close shot of a microplate

HTRF IP-One assay used for functional screening

Find out about the reliability of the IP-One HTRF® assay read with the PHERAstar microplate reader
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Legal remark

HTRF ® is a registered trademark of Cisbio.

Images partly taken from © fotolia.com

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