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Although the measured affinities are almost identical, the response amplitude increased more than three times
when p38α was labeled with the novel RED-tris-NTA 2nd Generation dye. Moreover, the signal-to-noise (S/N)
ratio increased by a factor of two using the new, TRIC-optimized dye (Figure 1 and Table 1).
Kit used to label target protein K
d
Amplitude S/N ratio
His-Tag Labeling Kit RED-tris-NTA (MO-L008) 8.66 ± 5.23 nM 2.19 11.63
His-Tag Labeling Kit RED-tris-NTA
2nd Generation (MO-L018)
9.15 ± 2.77 nM 7.71 22.70
References
1. Duhr, S., & Braun, D. (2006). Why molecules move along a
temperature gradient. Proc Natl Acad Sci U S A, 103(52),
19678– 19682.
*The amine reactive Protein Labeling Kit RED-NHS 2nd Generation
(MO-L011) and the cysteine reactive Protein Labeling Kit RED-Maleimide
2nd Generation (MO-L014) also offer TRIC-optimized dyes for MST
assays.
Conclusions
Based on a fluorophore specifically developed for
TRIC-detection, the novel Monolith His-Tag Labeling
Kit RED-tris-NTA 2nd Generation kit (MO-L018) yields
comparable affinities with the His-Tag Labeling Kit
RED-tris-NTA (MO-L008), but with increased binding
amplitudes and signal-to-noise ratios. The improved
performance of the 2nd Generation kit leads to a
higher detection success rate and reduces assay
optimization time, the two main advantages of
NanoTemper TRIC-optimized dyes*.
Table 1: Comparison of affinities, binding amplitudes and S/N ratios of the interaction between SB203580 and p38 labeled with RED-tris-NTA 2nd Generation
(MO-L018) or conventional RED-tris-NTA (MO-L008).
