Acetone pellets and protein insolubility

Michael Witty mw132 at mole.bio.cam.ac.uk
Mon May 21 06:36:49 EST 2001


Dear Paul,
          I would strongly advise you to phone the proteomics facility and
ask them what they prefer.  They may have some good suggestions - they
have done it all before!  Mike.

> Hi everyone,
>
> I am acetone precipitating proteins from the aqueous and detergent
> phases of a Triton X114 extraction. The problem is that the pellets if
> allowed to dry become hard like rocks and insoluble in normal 1D sample
> buffer for protein gels. I want to store the precipitated proteins for
> several weeks before I go to a specialised proteomics facility. I am
> worried if I freeze dry these pellets that they won't be of any use when
> i get to the proteomics facility. These are the ideas I have come up with:
>
> 1) Leave the proteins in acetone solution.
> - I've herd this can also make the pellet impossible to resuspend.
>
> 2) Try to resuspend the samples in PBS and then freeze dry or store at -70
> - I can't resuspend them in sample buffer because the proteomics place
> where i'm going has proprietary detergent solutions which they will
> resuspend them in.
> - I can't imagine the detergent phase proteins would go into a PBS solution.
>
> 3) Don't resuspend them in PBS, just layer it over the top of the
> pellets and store at -70.
>
> Please help or give any suggestions that you can.
>
> Thanks,
>
> Paul
>
>
> --
> Paul Cullen
> Bacterial Pathogenesis Research Group
> Monash University
> Australia
>




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