Steady-state FP recordings were performed
using a SpectraMax i3x
plate reader (Molecular Devices).61 (link),76 (link) All steady-state
FP measurements were conducted using a buffer that contained 20 mM
Tris–HCl (pH 7.5), 150 mM NaCl, 1 mM TCEP, 0.005% Tween 20,
and 96-well black untreated polystyrene microplates (Corning Inc).
Other details of steady-state FP measurements were previously reported.44 (link) 100 μL of each 20 nM labeled SET1Win peptide was added to individual wells at a final concentration
of 10 nM. The steady-state FP anisotropy was measured on the plates
after a 1 h incubation at room temperature in the dark. WDR5-dependent
dose-response data were averaged and then fitted using a four-parameter
logistic function to acquire the binding affinity (KD) for each interaction pairs.
using a SpectraMax i3x
plate reader (Molecular Devices).61 (link),76 (link) All steady-state
FP measurements were conducted using a buffer that contained 20 mM
Tris–HCl (pH 7.5), 150 mM NaCl, 1 mM TCEP, 0.005% Tween 20,
and 96-well black untreated polystyrene microplates (Corning Inc).
Other details of steady-state FP measurements were previously reported.44 (link) 100 μL of each 20 nM labeled SET1Win peptide was added to individual wells at a final concentration
of 10 nM. The steady-state FP anisotropy was measured on the plates
after a 1 h incubation at room temperature in the dark. WDR5-dependent
dose-response data were averaged and then fitted using a four-parameter
logistic function to acquire the binding affinity (KD) for each interaction pairs.
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