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Magnetic beads and elution


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#1 murtaugh

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Posted 08 January 2014 - 04:11 AM

Dear all,

 

I would like to purify low abundant protein from protein extract. I bind policlonal antibodies of this protein to magnetic beads from Invitrogen.

The protein extract consists 7M urea/2M thiourea/1% DTT. The elution buffer proposed by Invitrogen (cytric acid) is not working. Maybe solution with higher urea cc? Do you any ideas how to purify protein in such conditions?

 

Thanks in advance!

BR



#2 mdfenko

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Posted 08 January 2014 - 05:06 AM

are you sure your protein bound? (the antibody may be against the native rather than the denatured protein)

 

if so, sometimes high pH (10-11) is more effective for elution from antibodies than low pH (2.5). (don't forget to neutralize immediately after elution).


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