IUBio

Q: remove enzyme with antiserum?

Koen De Smet k.desmet at nospam.ic.ac.uk
Tue Mar 31 09:39:21 EST 1998


I am trying to remove selectively an enzyme from a crude extract using 
binding to an antiserum. In short, I hope to bind antibodies to the 
enzyme and then remove this by binding to beads coated with protein A 
and G.

But it doesn't work. My extract still has the same amount of enzyme 
activity.  I tried a number of approaches:

1/ first add the antiserum to the extract, incubate overnight at 4C, 
then add beads with protein A/G

2/ titrate antiserum in the above experiment

3/ first bind serum to beads for 4 hours, then wash and incubate with 
extract overnight at 4C.

Has anybody ever tried to do a similar experiment, have seen this 
published, or is anybody able to give me advise?

Thanks in advance
-- 
Koen De Smet
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