Proteins were first extracted from polyacrylamide gels using electroelution., Methods Mol Biol. 2007;355:353-63. Electroelution of intact proteins from SDS-PAGE gels and their subsequent MALDI-TOF MS analysis.Lei Z1, Anand A, Mysore KS, Sumner LW.

Electroelution of Proteins From Polyacrylamide Gels - by MJ Dunn - ‎2004

Hi Ravi thanks for the reply. Electroelution is what i'm actually doing and it works. I'm just interested if a possible ligand is co eluted after these treatments..

Whether or not the ligand remains bound during native PAGE and subsequent electroelution under nondenaturing conditions will depend upon the affinity of the complex. I would think that the ligand will be lost unless it has a Kd in the nM range.

The ligand can stay bound to the protein ideally  if it has a very strong affinity towards the protein. But I think it will dissociate from the protein during the down-streaming processes. 

Hello everyone, Im trying to elute my interest of protein from native page, but it is not working. im using elution buffer containing 50mM tris HCl, pH 8.2, 0.1M EDTA, 1mM DTT and 0.15M NaCl. i run the electrophorsis at 80volt for 3 hours.

can somebody verify, im doing in right way?

Hi Rimanpreet Kaur

How large is the protein of interest? I always electro eluted over night at 8°C 100V to have soft conditions. I also added 6-aminohexanoic acid as recommended by the authors in the nature methods bnpage paper to block proteinases. Do not use it if you plan trypsine digest for mass spectrometry as it inhibits trypsin. Large proteins are somewhat easier to electroelute. Check your success on a sds gel maybe with a non target protein as control.