Determination of dissociation constant of nusA-A1(Cys) for NusA by microscale thermophoresis


Overview
Abstract
The dissociation constant (Kd) of the interaction between nusA-A1(Cys) and NusA-GFP was measured by microscale thermophoresis. NusA-GFP was held at 100 nM while the concentration of nusA-A1(Cys) was varied from 10 uM to 0.305 nM while the migration of the fluorescent protein was measured upon local heating using a Monolith NT.115 Pico with 80% Laser Power, 20% LED power, at 25C.
Type
  • Biophysical Constant Measurement
Target
Scores
Outcome
Successful
Signal
Acceptable
Results
Figure

Image 

NusA affinity reagent

Legend 

Analysis of the interaction between nusA-A1(Cys) and NusA-GFP by microscale thermophoresis. Binding of nusA-A1(Cys) to NusA-GFP was quantified in PBS-T (0.05% Tween). NusA-GFP was held at 100 nM while the concentration of nusA-A1(Cys) was varied from 10 uM to 0.305 nM while the migration of the fluorescent protein was measured upon local heating using a Monolith NT.115 Pico with 80% Laser Power, 20% LED power, at 25C.