Figure 1

Diagram of the biochemical model screening assay. This diagram is taken from Wang et al. (2005) and illustrates the steps involved in the biochemical/immunoblotting assay used in this paper. The fusion protein GST-Q58-Htn (20 μg/ml) was mixed with thrombin (0.5 unit/μg protein) for 30 minutes and the mixture centrifuged to remove aggregated protein. The soluble protein was mixed with an oligonucleotide in a 96-well PCR plate and incubated for 24 hours at room temperature (RT). SDS was added to a final concentration of 2% and the mixture heated at 99°C for 5 minutes. Filtration through a 0.22 micron acetate cellulose membrane filter was followed by detection of aggregated Q58-Htn fragment by immunoblotting with an antibody (HP1) and ECL. Quantitation was carried out using an ImageQuant program. The blot displays both positive and negative results – positions lacking a black spot indicate that aggregation was inhibited by the oligonucleotide.