Protein elution from SDS-PAGE gel ??

Federici federici at icgm.cochin.inserm.fr
Wed Jul 3 05:47:46 EST 1996


In article (Dans l'article) <4r61f8$9mr at worak.kaist.ac.kr>, "Kernee H.
Chang" <Kernee at bioneer.kaist.ac.kr> wrote (écrivait) :

> Dear Netters
> 
> I'm trying to elute my glycoprotein from 1.5mm PAGE gel.
> 
> Could you tell me the best method for the case ?
> 
> And also the yield of the protein is important.
> 
> 
> Thank you very much....

electroelution combined with recuperation on hydroxyapatite is an
efficient method. This experiment is carried out on acrylamide gel
polymerized into tubes. First of all, prepare a tube containing a gel
filling about the third of the total volume; then add 0.5 ml of
hydroxyapatite washed by the buffer of electrophoresis (SDS, Na P...); cut
the gel containing the band  you intend to elute, into small pieces, and
put these pieces onto hydroxyapatite. fter electrophoresis, recuperate
hydroxyapatite and elute the protein using 0.5M NaP, SDS 0.2%, DTT.



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