How to show protein interaction?

Achim Recktenwald, PhD achim at ibex.ca
Thu Nov 28 07:54:28 EST 1996


Cornelius Krasel wrote:
> 
> Thorsten Schmidt (Thorsten.Schmidt at rz.ruhr-uni-bochum.de) wrote:
> > I have a protein and I just want to test if it forms dimers, trimers etc.?
> > (The protein was purified by HisTag-Chromatographie and is in solution
> > (20mM Tris, 0,5 M NaCl, 1 M Imidazol))
> > What is the simplest way to do this?
> 
> I'd try analytical gel filtration if you have the equipment (i.e. an
> FPLC/HPLC). Alternatively, you may try analytical ultracentrifugation
> (I have absolutely no experience with this) or maybe non-denaturing
> gel electrophoresis.
> 
> --Cornelius.
> 
> --
> /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
> /* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP3 */
> /* "Science is the game we play with God to find out what His rules are."  */


I would also try native polyacrylamide-gel-electrophoresis. If the bond
between the subunits is strong, you'll get one single band, the multimer
form; if its very weak, you'll only see a sharp band at the mol. weight
for the single subunit. But if the bond is neither nor than you get a
smear on your gel, starting at the point of the highest multimer down to
the lowest multimer, or even the single subunit. In the latter case the
multimer form is constantly decomposing during the electrophoresis.

Achim



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