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


Overview
Abstract
Analysis of the interaction between GFP-A1 and GFP by microscale thermophoresis. Note: a version of GFP-A1 with no C-terminal Cys was used in this experiment to eliminate possible disulfide interactions. Binding of GFP-A1 to GFP was quantified in PBS-T (0.05% Tween). GFP was held at 10 nM while the concentration of GFP-A1 was varied from 5 µM to 0.153 nM while the migration of the fluorescent protein was measured upon local heating using a Monolith NT.115 Pico with 20% Laser Power, 15% LED power, at 25C.
Type
  • Biophysical Constant Measurement
Target
Scores
Outcome
Successful
Signal
Acceptable
Cross Reactivity
Acceptable
Background
Acceptable
Dynamic Range
Acceptable
Results
Figure

Image 

Legend 

Analysis of the interaction between GFP-A1 and GFP by microscale thermophoresis. Binding of GFP-A1 to GFP was quantified in PBS-T (0.05% Tween). GFP was held at 10 nM while the concentration of GFP-A1 was varied from 5 µM to 0.153 nM while the migration of the fluorescent protein was measured upon local heating using a Monolith NT.115 Pico with 20% Laser Power, 15% LED power, at 25C.