Table 1.
Dissociating Agents | Protein recovered in soluble fraction ( % of total)b |
---|---|
Phosphate buffer (PBS) | 0.01 |
PBS – 1% Tween-20 | 0.012 |
PBS-1% Triton X-100 | 12 |
1 M NaCl | 0.001 |
2M MgCl2 | 0.001 |
8 M Urea | 62 |
1% SDS | 12 |
6 M Guanidium Hydrochloride | 68 |
α-crystallin and αA66-80 peptide (2:1) ratio (W/W) were incubated at 37°C for 18 h. After incubation, the samples were centrifuged (10,000 g, 10 min) and the aggregates were resuspended in different dissociating agents and placed in roller shaker at 37°C for 1 h. The samples were again centrifuged at 8000 rpm for 10 min. Protein content of the supernate was estimated by Bio-Rad protein assay. Protein content of the sample at 0 min before incubation was taken as 100%.
Bold-faced percentages indicate the maximum solubility of the aggregate in 8 M urea and 6 M guanidium hydrochloride