HTRF technology requires simple yet specific detection conditions. The HTRF signal is detected at two different wavelengths (typically 620 nm and 665 nm) which are used to calculate the fluorescence ratio and compensate for compound interference and sample quenching. Emissions are also detected using a time-resolved module which enables the introduction of a delay between the excitation pulse and the time-gated measurement window, which greatly diminishes background fluorescence detection.
Optical devices such as excitation and emission filters, monochromators and dichroic mirrors, as well as energy source and photomultiplier tubes, constitute critical equipment for measuring HTRF output. Instrument set-up has a significant impact on detection efficiency, in particular the measurement window and time delay, excitation parameters (e.g. number of flashes delivered), gain adjustment, and reader head positioning with respect to the well. The reader control kit can be used to assess the performance of compatible readers to read HTRF technology using either Europium or Terbium cryptate chemistry.
In order to broaden HTRF readout capabilities for use in the field of life sciences, Cisbio Bioassays has initiated partnerships with several detection system manufacturers. This collaboration has led to reader certification, indicated by an HTRF sticker. This label guarantees that the reader meets the physical specifications for optimal HTRF readout.
The following instruments have already been certified for HTRF readout:
|Infinite F200 PRO
Infinite M1000 PRO
|Mithras LB 940
You may also check the compatibility of your equipment by contacting your local Cisbio Bioassays sales and application support team.
The HTRF reader control kit is designed to calibrate all HTRF-compatible readers. This immunoassay-based kit includes all the necessary reagents (conjugates and calibrators) as well as buffers and one 96 half-well microplate. The plate map includes a low and a high level calibrator, conjugate blanks, and a specific control to assess the counting rate at 620 nm (i.e. Eu3+ Cryptate emission level). The assay may be run in two different incubation modes: 3 hours or overnight at room temperature. The same plate (i.e. experiment) may be used to check several instruments.
This kit may also be used to validate readers following their upgrade to HTRF compatibility. It provides an easy and straightforward way to check the overall performance of a reader and may facilitate the identification of potential failures.
|Description||Quantity||Cat no.||Product Insert||MSDS|
|HTRF reader control kit||1 kit||62RCLPEA|