NEED HELP with non-denaturing IEF

[Jeffrey Yuziuk]

Dear Aye,

I am also a graduate student. I have run different kinds of IEF gels. 
Some of the considerations I have found to be important for viewing 
isozymes are: carefully check the salt concentration in your samples. 
10mM NaPhos should be OK, but not too much higher; the time elapsed 
til you visualize activity is important, due to diffusion, both of your 
enzyme bands, and the substrate. If you cut the gel in half, and stain 
one side for activity, and the other for protein, do you see focused 
bands in the protein-stained side for your samples? 
Hope this helps a little...

Jeff