Dear Sir. Concerning your issue about the less value for control in BSA denaturation assay. aluable information on the initial stage of BSA aggregation can be obtained from the portion of the aggregated protein versus the portion of the denatured protein plots. The portion of the aggregated protein (γagg) was calculated as (1 –γnon-agg) and the portion of the denatured protein (γden) was calculated as (1 –γnat). Fig 5 shows the γagg vs γden plots obtained at 60°C, 65°C, 70°C and 80°C. Particular attention has been given to Fig 5B (65°C). At this temperature the relationship between γagg and γden is linear. This means that unfolded protein rapidly aggregates without accumulation in the solution (the rate of aggregation significantly exceeds the rate of denaturation). The length cut off on the vertical line passing through γden = 1 by the linear dependence of γagg on γden gives the portion of the unfolded protein participating in fast aggregation stage (γUhr, the portion of the highly reactive unfolded protein). At 65°C γUhr is equal to 0.51±0.03 (Table 1). The remainder of the protein (1 –γUhr) is involved in the aggregation process with the relatively low rate and can be called the low reactive unfolded protein (γUlr). At 65°C γUlr is equal to 0.49. Taking into account that at this temperature γnon-agg,lim = 0.35, we can conclude that the portion of the low reactive unfolded form involved in the formation of large-sized aggregates (γUlr,agg) is equal to 1 –γUhr−γnon-agg,lim = 0.14 . For more details, I think the following below links may help you in your analysis:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4839713/pdf/pone.0153495.pdf

Thanks

Hi Sunim, have you anticipated the free -SH in serum albumin?

No not at all sir.