Hi Anamika,
As it was said before, a cocktail of protease inhibitors should be
fine. But let`s say you can not overcome the problem. It is not specified
if you are doing this dialysis to cleave or not your His-tagged protein but
even this you could try getting samples in 1 hour intervals then you could
monitor the stability (and/or cleavage) of your protein. Another thing is
if you are doing this at room temperature or at 4C.
Best regards,
Everton.
On Thu, Oct 26, 2017 at 5:11 PM, Smith Liu <[log in to unmask]> wrote:
> suppose protease is the issue, then avoid overnight dialysis
>
>
> 发自网易邮箱大师
>
> 在2017年10月26日 22:18，Anamika Singh <[log in to unmask]> 写道：
> Dear All,
>
> I am purifying the His-tagged proteins ( 21Kda) using buffer 20mM Tris,
> 150mM Nacl, and 5mM MgCl2 with 500mM imidazole. Earlier I was facing the
> precipitation problem during dialysis but with the addition of 50mM L-Arg
> somehow managed to overcome the precipitation issue. But this time I have
> seen after overnight dialysis there are degradation products on SDS page. I
> have used protease inhibitor (PMSF) in my sonication buffer.
>
> Please suggest me to overcome this problem.
>
> Thanks
> --
> Anamika
>
>
>
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>
--
Éverton Dias D'Andréa
