SDS-PAGE gel problem

skoufos at skoufos at
Wed Dec 7 14:44:01 EST 1994

In article <3c3arl$5nu at>, ez027154 at (Kitty Kit Chui) writes:
>In the past, other people in my lab were able to resolve the 
>phosphorylated and unphosphorylated forms of the protein in SDS-PAGE gel 
>with 5% for running and 3% for stacking.  The ratio of acrylamide : bis 
>is 30%-0.8%.  The phosphorylated form usually runs at a much higher 
>molecular weight than the unphosphorylated one.  However, my gel kept 
>giving only one band.  


>Kitty Chui
>Section of Molecular Biology and Biochemistry
>UC Davis

One of the things you may want to check is your bis.  Its quality can change
with time and with incorrect storage.  Another thought is to try 2D gels that
will probably give you better separation in the 2nd dimension due to the 
charge change in your protein. I hope this is of help.


Emmanuel Skoufos, Ph.D.
Insect Molecular Biology Group
Institute of Molecular Biology and Biotechnology
P. O. Box 1527
Heraklion, Crete GR71110

More information about the Methods mailing list