Gel-elution

Ron Tate rtate at bmb-fs1.biochem.okstate.edu
Mon Nov 27 14:29:47 EST 1995


fester at mpimg-berlin-dahlem.mpg.de wrote:
>Hello Netters,
>
>I would like to elute protein complexes (mitochondrial respiratory
>complex III) from Blue Native Gels (about 7% acrylamide) and have
>no experience in doing so. I know electroelution is certainly the best
>way to go, unfortunately I don't have the necessary equipment.
>Is it possible to elute the proteins under native conditions or have
>I to denature them? Is there some blotting-membrane where proteins can 
>be eluted from after electro-transfer from the gel?
>Is it possible at all to elute proteins in another way than electroelution
>from a PAGE? 
>
>Many thanks in advance 
>
>Thomas Fester
>---------------------------------------------------------------------------------
>fester at mpimg-berlin-dahlem.mpg.de
>Ihnestrasse 63
>D-14195 Berlin
>Germany
>Institut fuer Genbiologische Forschung Berlin
>

Thomas,
 You can probably elute native proteins but recovery will probably be much lower than using 
SDS.  I have seen reference to desorption of protein from PVDF in a book about protein 
preparation for microsequencing edited by Paul Matsudaira.  Maybe another low-tech way to 
attempt this is using Amicon's micro filter/concentrator units.  They now have a nebulizer 
that turns gel pieces into a slurry in the centrifuge, they also have a protocol for getting 
proteins out of PAG.  I have not tried them yet but I have some ordered for that purpose.
Hope this helps.
Happy Hunting
Ron Tate, OSU BMB




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