2 replies to this topic

[perionix]

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

[laurequillo]

perionix, on 20 August 2010 - 03:09 AM, 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?

[perionix]

Elution: peptide (Sigma): 200ug/ml- 30 min at 4°C..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