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Problems with affinity purification with M2 beads


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#1 Mylando

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Posted 29 December 2009 - 08:37 AM

Dear fellow scientists,


I'm doing the affinity purification for flag-tagged proteins with M2 beads from Hela nuclear extracts. But I found this protein partially precipitated in the beads and can not be eluted completely. Do you have explanations for this phenomenon? What can I do to improve this?


kind regards,


Mylando

Edited by Mylando, 29 December 2009 - 08:41 AM.


#2 miBunny

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Posted 30 December 2009 - 07:19 PM

What kind of protein are you working with? Why do you think the protien partially precipiated on to the beads? Can you provide some more information on when you think your protein crashed out?

#3 Mylando

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Posted 31 December 2009 - 07:58 AM

Hi, thank you for replying. I am working with one of the linker histones. Because the proteins precipiated on the beads can not be eluted by 0.1M glycin at PH 3.0, but can only be boiled down. Could you give some suggestions for avoiding this precipitation?

Edited by Mylando, 31 December 2009 - 08:03 AM.





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