TECHNICAL NOTE
Protein Labeling
Improved quantitation of biomolecular interactions by MST using the His-Tag Labeling
Kit RED-tris-NTA 2nd Generation
Introduction
With the goal of improving the quality and robustness of MicroScale
Thermophoresis (MST) measurements, we enhanced our protein
labeling kits by selecting fluorophores that are more sensitive to
temperature changes (Temperature Related Intensity Change or
TRIC) and are therefore superior reporters of binding events.
MST is a biophysical technique that measures the strength of
the interaction between two molecules by detecting variations
in fluorescence signal measured from one of the binding partners
as a result of an IR-laser induced temperature change. The range of
the variation in the fluorescence signal correlates with the binding
of a ligand to the fluorescent target
1
. TRIC is an effect where
the fluorescence intensity of a fluorophore is dependent upon
temperature changes and binding interactions. Thermophoresis is
the quantifiable movement of fluorescent molecules along a
temperature gradient. Both TRIC and thermophoresis contribute
to the variation of fluorescence measured by MST.
This TRIC-optimized generation of dyes boosts the sensitivity
of MST measurements and yields vastly improved binding
amplitudes and signal-to-noise ratios. Incorporated into the
2nd Generation kits, the RED-tris-NTA 2nd Generation dye,
Ivana Bekić, Matthias Molnar and Nuska Tschammer
NanoTemper Technologies GmbH, Floessergasse 4, 81369 Munich, Germany
