Acetone pellets and protein insolubility

Paul Cullen Paul.Cullen at med.monash.edu.au
Mon May 21 03:00:46 EST 2001


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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