The polybasic head indicative of your protein to have some possible
membrane association? Try a detergent in your buffers might help. Also some
of the polybasic head for protein need lipid association to be happy (PI
and PCh)?
Padayatti
On Fri, Aug 23, 2013 at 6:27 PM, Jahan Alikhajeh wrote:
>
i am not sure this would work as his protein seems to be degraded by the n
end degradation pathway. i feel like it almost needs to be expressed as a
fusion protein with some stabilizing sequence
On Fri, Aug 23, 2013 at 10:08 PM, Roger Rowlett wrote:
> Why not adopt a classical purification strat
Why not adopt a classical purification strategy? IEX-HIC-GEC. His-tag not
required. With your protein strongly basic, anion exchange seems like a
likely first step. After IEX, hydrophobic interaction or salt precipitation
followed by gel exclusion is normally enough for well-expressed proteins.
Ro
Dear Friends,
I have been trying to purify a protein (27 kDa) which has a sumo plus 6 His-tag
at N-ter giving totaly a 45 kDa protein.
This protein does not express without sumo tag and has a poly basic tail (Arg
and Lys) at its N-ter. It does polymerize at acidic pHs.
When I tried to purify it
