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M2 Flag purification - (Aug/20/2010 )

Hi,

I've been trying to elute my protein from the M2Flag beads but with no luck.
I tried low pH, different concentration of peptide...and still the same result...the protein comes out only after boiling the beads.

How can I improve the elution?

Thanks

-perionix-

perionix on Fri Aug 20 11:09:29 2010 said:


Hi,

I've been trying to elute my protein from the M2Flag beads but with no luck.
I tried low pH, different concentration of peptide...and still the same result...the protein comes out only after boiling the beads.

How can I improve the elution?

Thanks

How did you do the elution, and what concentrations of flag-peptide did you use? it is the 3xFlag or Flag?

-laurequillo-

Elution: peptide (Sigma): 200ug/ml- 30 min at 4C..one elution....
Than I tried to increase the elution numbers...and even the concentration up to 1mgr/ml....the result was the same....
I have 1xFLAG

-perionix-