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


Overview
Abstract
The dissociation constant (Kd) of the interaction between AVD-A1(Cys) and Avidin was measured by microscale thermophoresis. AVD-A1(Cys) was conjugated to an AlexaFluor and held at 5 nM while the concentration of Avidin was varied from 5 µM to 0.153 nM, and the fluorescence measured upon local heating using a Monolith NT.115 Pico with 40% Laser Power, 30% LED power, at 25C.
Type
  • Biophysical Constant Measurement
Target
Scores
Outcome
Successful
Signal
Acceptable
Results
Figure

Image 

MST Dissociation Constant nanoCLAMP AVD-A1 and Avidin

Legend 

The dissociation constant (Kd) of the interaction between AVD-A1(Cys) and Avidin was measured by microscale thermophoresis. AVD-A1(Cys) was conjugated to an Alexafluor 568 and held at 5 nM while the concentration of Avidin was varied from 5 µM to 0.153 nM, and the fluorescence measured upon local heating using a Monolith NT.115 Pico with 40% Laser Power, 30% LED power, at 25°C