Supplementary Figure 3: Bead-based assays confirm bulk Edman sequencing of fluorescently labeled amino acids.
From: Highly parallel single-molecule identification of proteins in zeptomole-scale mixtures
(A) Specific binding of TMR to functionalized Tentagel beads occurs at the periphery and density can be measured by image processing. Peripheral bead fluorescence intensities were calculated by computing the area under the fluorescence intensity radial distribution normalized relative to a negative control beads bound non-specifically with free TMR lacking the NHS group to control for background fluorescence. (B) Edman degradation can be used to determine the positional information of the fluorescently labeled lysine residues of synthetic peptides using bulk fluorescence measurements. Bar charts indicate the normalized average of the fluorescence intensity per bead +/- s.d. across image fields. Raw fluorescence intensities and field counts are reported in Supplementary File 1.
