Figure 3. SDS-PAGE high throughput screen.
Example gels from the SDS-PAGE based high throughput screen. A: As described in the text, a 12x5 section of a 96-well plate contains one library member per well in SDS gel loading buffer. In the sixth row, a portion of each well above is added to give a mixture of five peptides. B: Sixty peptides are analyzed on a single gel in 12 lanes, with 5 peptides per lane. About 1 µg of Neu/V664E is analyzed in the 8th lane. Most library members migrate as monomers. C: When a putative dimer is identified (e.g. lane 11 in panel B), a second gel is run with the remainder of the five single peptide solutions to identify the dimeric peptide. In this secondary gel, a sample of Neu/V664E is included in lane 3. D: Most gels analyzed contained no dimers, as shown by this typical example.