Visualising 200-400+ kDa proteins by gel electrophoresis - (Feb/05/2007 )
I have a protein which is 200kDA and is found to oligomerise so I need to be able to see band separation at 400, 600 kDA etc. What percentage gel would I need to use? I have heard that you can carry out this kind of experiment using an agarose gel and by buying kits but I would prefer to make the running buffers myself. Thank you.
use a gradient gel; for separating the largest known proteins, titin (~1000 kDa) and its variants, often starts with 2% to 16%; if you plan to run a native gel, you should know the pI of your protein to choose the right buffer system...
to separate high mw proteins we used a 3.5-5% acrylamide gel with a 0-8M urea gradient (to sharpen the bands). we use the neville buffer system for sds-page but this should work with the laemmli buffer system.
Thanks very much I shall take this on board!