Magnetic anti-flag beads

From: Harman, Christine
Date: 17 November 2011 19:29

Hi all,

Sorry for the off-topic question, but I wanted to find out anyone's experience with using magnetic anti-flag beads (Sigma).  I have been having a problem with the anti-flag antibody leaching from the beads during low pH elution ( which shouldn't be happening I am thinking) and was wondering if anyone has had a similar experience.  I have talked to Sigma about this and they sent me another lot of beads which also appears to be leaching antibody during incubation with 0.1M glycine pH 2.7 (normal elution described in protocol except at pH 3.0).  I am not using the beads for purification, but will be using them for protein interactions studies so sorry about the off-topic question.  Given that you all are a very smart community I thought I would throw this one out to you.

 

Thanks for any brain waves on this,

 

Christine

 

 

 

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From: Ho Leung Ng


    If it's the M1 antibody, you can elute with EDTA + Flag peptide.
You could also try a little higher pH like 3.5.


Ho Leung Ng
University of Hawaii at Manoa
Assistant Professor, Department of Chemistry