Native polyacrylamide gel question

[Mike Marr]

What is th pI of the protein if the protein is positively charged at the
pH of your running buffers it will not migrate towards the (+)
electrode. You can play around with the pH of the buffers to try to give
the protein a negative charge.

Mike Marr
Section of Biochemistry
Cornell University.

ccc6 at Lehigh.EDU wrote:
> 
> Hi,
> 
> A colleague of mine is having trouble with running native polyacrylamide gels
> of her protein.  On denaturing gels, there's been no problem seeing her
> isolated protein clearly.  However, it seems with the native gels, the protein
> is not even entering the gels.  She's tried lowering the percentage of
> polyacrylamide as well as running the gels for longer periods of time, but the
> protein does not seem to be entering the gel.  As I'm posting this for someone
> else, I don't have all the details of what she's done right in front of me,
> but I could find out easily.  Has anyone had a similar experience or been able
> to fix a similar problem??
> 
> Any suggestions would be greatly appreciated
> 
> Christine