Protein-Protein Interaction by Native Gel

Greg Pankhurst g.pankhurst at spamblock.cfi.unsw.edu.spamblock.au
Sun Feb 15 22:31:14 EST 2004


<lcrowley at hsc.usf.edu> wrote:

> I am currently working on a project showing the enhanced binding of two
> proteins via site directed mutagenesis and would like to visualize this
> by Native Gel. I have made a poor attempt at this using a protocol from
> Sigma-Aldrich on a 12.5% pre-cast Bio-rad gel and my proteins completely
> separated on the gel. Are there any protocols out there that I could use
> that would help me with this, perhaps by lowering the ionic strength of
> the running and/or sample buffers.

Why not try crosslinking the complex together and run SDS gels ?? My
guess is it'll give you much cleaner data than native gels.

Pierce sell a whole bunch of crosslinking reagents with different
chemistries.

Greg



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