NEED HELP with non-denaturing IEF
jayuziuk at rs4.tcs.tulane.edu
Tue Sep 26 14:31:18 EST 1995
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...
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