Make my own resin?

Wolfgang Schechinger novalidaddress at nurfuerspam.de
Wed May 3 17:17:02 EST 2006


Hi,

I'd recommend a crude purification first: Ammonium sulfate
fractionation, followed by DEAE chromatography or whatever you want to
get rid of most unwanted stuff (eg proteases!) first. Then, you either
may load the antibody on some protein A or G column and use it for
purification (but your AB probably will go off during the elution, too
and then all your AB will be lost, for a second batch you'll need a new
load).
So, better bind your AB covalently to a reactive resin (these tiny
little pre-packed NHS activated agarose columns from
Amersham/Pharmacia/GE Healthcare or what their current name is are
graet for this, but CNBr activated agarose and self packed column will
do as well. If you want/need to polish your stuff afterwards, that will
depend on your contaminants. So gel filtration, T7 affinity column, HIC
or something special else might still be neccessary.

HTH

Wolfgang Schechinger
Endocrine Research Lab
University Hospital "Bergmannsheil"
Bochum, Germany

PS please reply to the group, the email address above ends in autotrash



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