Thank you for your response. The problem I have is with binding the protein
on the HIC column (Phenyl). The urea is having adverse effects on binding.
Is there a way to circumvent that, bearing in mind that the protein
self-aggregates easily? Sorry I haven't been clear in my message.
H.V.Taylor wrote in message <7eN2OnDKOI+OfGeMjpIP84QZNn4a at 4ax.com>...
>>What type of HIC are you using? If it's C4-, C8- or phenyl- you could
>try adding organic solvents to elute (your protein is denatured
>anyway). I'd start with glycerol or ethylene glycol as they will more
>likely be compatible with >5M urea than other solvents. However, the
>self-aggregation of your protein may be suppressed by glycerol or
>ethylene glycol as well. As usual, it's a question of trial and