want to purify my proten!!!!!!!!

Yong Jiang yjiang at ucsd.edu
Fri Feb 18 15:34:15 EST 2000


Hi,
I think there are at least two ways to purify protein from PAGE gel, the
recover efficiency is dependent on your gel conc., protein size, and stain
or not. One is electroelution, the other is mechemical elution. For
electroelution, you can cut the gel slice containing the target protein(if
you did not stain it, you can use 0.25M KCl dye the SDS-PAGE gel 10 min,
destain with ddH2O for another 5~10 min, the bands should be seen against a
black background), put it into a dialysis tube, electrophoresis 40~60 min,
before stop it, changing the current direction for 30s is necessary, then
collect the buffer in the dialysis tube, concentrate it or dialysis against
water before concentration, it is optional. For mechamical elution, you can
use Amicon Co's column( I can not remember it's name exactly, you can querry
the company for that), it can squeeze the gel slice and elute the protein
from the gel. I hope this is helpful.
Best Regards!
Yong
Mike Zharkovsky <mike_z at ut.ee> wrote in message
news:01bf7a07$2a377320$670e28c1 at imlab5.ut.ee...
> Hi
> Could anybody write me how I can purify my protein from Acrylamid gel
after
> electrophoresis?
> thank you
>
> Mike Zharkovsky
> mike_z at ut.ee






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