NanoTemper Monolith instruments have gained enormous popularity for measuring molecular interactions both in academia and industry. The underlying technology has been extensively reviewed along with its assumptions, limitations, and applications (Scheuermann et al., Anal Biochem 496:79-93, 2016). Several assumptions about the technique such as the extent of thermal deviations generated by the infrared laser and the thermophoretic foundation of the measured signal have been revised during the last decade. We present here in this letter the experience gathered in scientific service facilities about this technique and make scientists aware of possible pitfalls with the intention to promote knowledge and good practice throughout the scientific community.
Keywords: Interaction; K D; MST; TRIC; Thermophoresis.