Effect of buffer on K<sub>D</sub> of SMAC peptide binding to BIR domains.
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Abstract
<p>All assays contained TCEP at 1 mM, 0.005% Tween 20 and SMAC-rhodamine at 20 nM. The buffers used were PBS @ pH 7.4, 25 mM HEPES @ pH 7.5, 25 mM HEPES @ pH 7.5 with 20 mM β-glycerol phosphate, 10 mM Potassium Phosphate @ pH 7.4, or 50 mM TRIS @ pH 7.5. Proteins were diluted into 25 mM HEPES @ pH 7.5 with 1 mM TCEP. FPA data were collected on the Analyst at 0, 30 and 60 min. Time overlays are plotted in the figure. K<sub>D</sub>s were determined in Prism.</p
