GFP on SDS-GEL
rdudley+ at pitt.edu
Wed Sep 23 09:20:17 EST 1998
Jan-Peter Siebrasse wrote:
> Rich Dudley wrote:
> > On SDS-PAGE, no. I'm not sure about native PAGE, but probably.
> > rich
> From my own experience, yes. Most of the GFP/GFP fusion protein will not
> be denatured as long the sample will not be heated or your gel is
> running too hot. And you have to load your sample immediatly after
> mixing them with sample buffer. After fixation fluorescence will be
> lost, of course. I even noticed a clearly visible fluorescent band after
> transferring GFP/GFP fusionproteins onto a NC membrane for Western
> BTW, the non-denatured GFP runs at an apparently lower molecular weight
> (ca. 8kDa "smaller" than the denatured protein). Most likely due to the
> more compact structure of the native protein I think.
> Jan-Peter Siebrasse
Just out of curiosity, how do you visualize the green--with a
I usually include boiling in SDS-PAGE buffer as part of my protocol. I'll
have to try this!
--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley+ at pitt.edu)
Research Specialist V
Dept. of Cell Biology and Physiology
University of Pittsburgh
---> search BIONET archives at http://www.bio.net <---
-------------- next part --------------
A non-text attachment was scrubbed...
Size: 463 bytes
Desc: Card for Dudley, Richard
Url : http://iubio.bio.indiana.edu/bionet/mm/methods/attachments/19980923/08bcc3fc/vcard.bin
More information about the Methods