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TAP Tagged Affinity Purification + Sticking to IgG-seph

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#1 Geert



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Posted 27 September 2004 - 05:02 AM


We are using as an initial step to capture our protein complexes(from plant cell cultures) Ig-G sepharose6 Fast Flow(Amersham Biosciences).
The tagged protein we're trying to capture has a TEV-cleaving site.
And we believe that even after TEV-cleavage still a lot of our protein sticks to the affinity-resin.
The evidence for this is after cleaving by boiling the resin in SDS-sample buffer something like 50% is found back.
Are you encountering the same problems?
And do you have some suggestions to solve this problem?

Yours sincerely,

Geert Persiau
VIB-Plant Systems Biology


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