How:
1. measure total protein before refolding (eg by absorbance at 280)
2. Purify refolded fraction by gel filtration in FPLC
3. measure total protein after refolding
4. calculate efficiency ...
Why?
I don't see the reason to know the efficiency. If after refolding I
would get enough in a defined molecular species,
soluble in a non-denaturing buffer, I would be happy. 'Enough' depends
on what you want to do with it.
Some refolding protocols have efficiencies of just 2-5%, but this is
enough ...
A.
On Feb 10, 2011, at 10:45, megha goyal wrote:
Hello,
Can anyone let me know how to calculate solubilisation and refolding
efficiency. We perform solubilisation and then refolding and check
by HPLC if refolding is completed or not [single peak on HPLC]. how
do i determine % efficiency of refolding. kindly guide me.
regards,
megha
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Anastassis (Tassos) Perrakis, Principal Investigator / Staff Member
Department of Biochemistry (B8)
Netherlands Cancer Institute,
Dept. B8, 1066 CX Amsterdam, The Netherlands
Tel: +31 20 512 1951 Fax: +31 20 512 1954 Mobile / SMS: +31 6 28 597791
