Sucrose gradient - (Aug/27/2001 )

I need to run a sucrose gradient with recombinant proteins. I cannot find a good protocol describing the general conditions (density, salt concentration, run time, speed etc)related to the molecular weights of my proteins. One protein has a MW of 14kD, and the other of 70kD. I am hoping to see if these proteins interact to form multi-subunit conplexes of higher molecular weight. Can anyone help?Thank you.

Hi, Avak

The sucrose gradient it is not a good method for protein separation (especially with a small MW). You will lost a lot of time for installation and irreproducible resulst. As I think, the best method for solving your problem is the ion-exange or gel-permeation chromatografy. These methods are highly reproduceble and simple for use.Good luck,Oleg Klychnikov, PhD.