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M2 Flag purification


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#1 perionix

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Posted 20 August 2010 - 03:09 AM

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

#2 laurequillo

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Posted 20 August 2010 - 03:13 AM

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?
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#3 perionix

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Posted 20 August 2010 - 03:32 AM

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




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