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Affinity purification of antibody against MBP-fusion

Affinity purification of antibody against MBP-fusion - Protocols and Methods Forum

Affinity purification of antibody against MBP-fusion - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 06-04-2008, 09:31 PM
John Leonard
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Default Affinity purification of antibody against MBP-fusion



I have an antibody against a protein I'm studying, and I'd like to purify it
from the serum to see if I can improve its specificity for my protein in
Western, EMSA, ChIP, etc. I have a vector for an MBP fusion of my protein
(which I know the antibody binds very well), and I was hoping to use that to
affinity purify my antibody. Ideally, I would like to express the bacterial
fusion, run it over an amylose column, then use the column-bound MBP fusion
to affinity purify my antibody. I have seen similar protocols using His- or
GST-tagged proteins, but have not found a protocol for MBP fusions.

If anyone has a protocol for something like this, or has another idea for
how I can purify my antibody based on specific ligand interaction, I'd
greatly appreciate it!
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