[Protein-analysis] 2D gel lossing high molecular weights

[Isabelle Holdridge]

Hello,
 
I have been running 2D gels for soy bean total soluble proteins and I am consistently losing the higher molecular weight proteins.  These same proteins are visible on the 1D gels.  I have tried a variety of changes in my protocol, such as, changes with the running buffers, IPG buffers and detergents, and ph gradients, but all to no avail.  The lower molecular weights on the 2D gel are visible and appear to be nicely focused.  I am not getting much background staining or streaking.  Any suggestion would be appreciated.
 
Thanks
Isabelle