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THUNDER™ TR-FRET with BMG LABTECH’s PHERAstar® FSX: a powerful approach for high sensitivity cytokine quantification


As part of our ongoing strategic collaboration with BMG LABTECH, we have recently evaluated the performance of a panel of five THUNDER™ TR-FRET human cytokine assays on the top-of-the-line PHERAstar® FSX multi-mode microplate reader. The results, summarized in the Application Note 365, showed a superb assay performance of our assays on this HTS instrument.

Figure 1: THUNDER™ Human IL-1β standard curve.

Figure 2: A linear plot of the low concentration part of the THUNDER™ Human IL-1β standard curve.


Table 1: Performance of THUNDER™ human cytokine assays on the PHERAstar FSX and the competing HTS reader.

Analyte Assay parameter PHERAstar FSX Competitor
IFNγ LOD (pg/mL) 6 24
LLOQ (pg/mL) 33 121
S/B 15 9
IL-1β LOD (pg/mL) 1 26
LLOQ (pg/mL) 7 141
S/B 23 17
IL-12 LOD (pg/mL) 3 20
LLOQ (pg/mL) 19 98
S/B 22 15
TNFα LOD (pg/mL) 3 13
LLOQ (pg/mL) 41 62
S/B 57 45
CCL2 LOD (pg/mL) 3 13
LLOQ (pg/mL) 17 61
S/B 17 15



  • The THUNDER™ human IFN-gamma, IL1-beta, IL-12, CCL2, and TNF-alpha assay kits were tested in a 384-well format using the standardized protocols.
  • Assays were read on the PHERAstar FSX equipped with a 377 nm laser excitation source.
  • For comparison, plates were also read on a competing HTS microplate reader equipped for TRF laser excitation.
  • The limit of detection (LOD) and lowest limit of quantification (LLOQ) were calculated by adding 2 or 10 standard deviations (SD), respectively, to the mean background counts (zero standard; 12 replicate wells).
  • For all assays tested, the PHERAstar FSX exhibited broader dynamic ranges and higher sensitivity compared to the competitor and our internal data, decreasing the LOD to single digit pg/mL.
  • These data demonstrate that the PHERAstar FSX is particularly advantageous for performing quantitative THUNDER™ cytokine assays.
  • PHERAstar® FSX is therefore certified for use with THUNDER™ TR-FRET.