Handling for Monolith NT.115 Series and NT.LabelFree.
One of the binding partners is labeled using standard fluorescent labeling protocols. Blue, green or red dyes and all coupling chemistries are compatible with our technology. The MST signal is also independent of the position of the fluorescence dye on the molecule. NanoTemper provides dyes optimized for protein compatibility and MST analysis. When using the Monolith NT.LabelFree, no labeling is necessary.
Preparing the Interaction Experiment
A titration series is prepared, where the concentration of the fluorescent binding partner is kept constant and the concentration of the unlabeled (i.e. non fluorescent) molecule is varied.
C. Transfer into the Capillary
After an incubation time sufficient for the reaction to reach equilibrium (e.g. 5 min.), the samples are transferred into NanoTemper capillaries or capillary chips. The single capillaries are placed on the sample tray which accommodate up to 16 capillaries, whereas one capillary chip can be loaded with 24 samples.
D. Measurement and Data Analysis
Insert the capillary tray or the capillary chip into the instrument. Close the door of the instrument by using the touch display of the Monolith series instrument. Simply start the software and the instrument automatically recognizes the presence of properly filled capillaries on the sample holder. It automatically measures the thermophoresis signal of each sample and calculates the dissociation coefficient. The analysis software highlights protein aggregation events, false positives and discriminates epitope/binding sites.
Movie about handling for Monolith NT.115 Series and NT.LabelFree.