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Excellent performance of THUNDER™ phospho-protein and cytokine TR-FRET assays on BMG LABTECH’s CLARIOstar® Plus


In the framework of our ongoing strategic collaboration with BMG LABTECH, we have evaluated the performance of a panel of three THUNDER™ TR-FRET phospho-proteins assays and five human cytokine assays on the flexible and powerful CLARIOstar® Plus multi-mode microplate reader. The results, summarized in the Application Note 361 and in a tutorial showed an excellent assay performance of our assays on this very sensitive instrument.


Fig. 2: THUNDER™ Phospho-ERK1/2 (T202/Y204) assay (4 hours incubation).


Fig. 3: THUNDER™ Phospho-p38αβγ (T180/Y182) assay (overnight incubation).


Fig. 4: THUNDER™ Phospho-STAT3 (Y705) assay (overnight incubation).


Table 1: Performance of THUNDER™ human cytokine assays on the CLARIOstar Plus (xenon flash lamp) and the competing HTS reader (TRF laser).

Analyte Assay parameter CLARIOstar Plus Competitor
IFNγ LOD (pg/mL) 4 10
LLOQ (pg/mL) 22 70
S/B 11 9
IL-1β LOD (pg/mL) 7 36
LLOQ (pg/mL) 42 164
S/B 23 18
IL-12 LOD (pg/mL) 4 38
LLOQ (pg/mL) 25 179
S/B 19 15
TNFα LOD (pg/mL) 3 41
LLOQ (pg/mL) 28 211
S/B 58 42
CCL2 LOD (pg/mL) 9 44
LLOQ (pg/mL) 40 172
S/B 11 9



  • Human IFN-gamma, IL1-beta, IL-12, CCL2, and TNF-alpha assay kits were tested in a 384-well format using the standardized protocols.
  • Phospho-ERK1/2 (T202/Y204), Phospho-p38αβγ1/2 (T180/Y182), and Phospho-STAT3 (Y705) assay kits were tested in a 384-well format using the 2-plate (transfer) protocol.
  • Assays were read on the CLARIOstar Plus equipped with a high energy xenon flash lamp excitation source.
  • For comparison, plates were also read on a competing HTS microplate reader equipped for TRF laser excitation.
  • For cytokine assays, 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 CLARIOstar Plus exhibited broader dynamic ranges, higher sensitivity and S/B ratios compared to the competitor and our internal data, decreasing the LOD of cytokines to single digit pg/mL.
  • The combination of simple, rapid, and robust yet affordable THUNDER™ TR-FRET assays with the highly sensitive CLARIOstar Plus provides a powerful approach for accelerating the workflow of drug discovery and life science research.
  • CLARIOstar® Plus is therefore certified for use with THUNDER™ TR-FRET.


THUNDER certification