Help on Extracellular matrix analisys

James Bassuk bassuk at u.washington.edu
Thu Mar 26 00:59:33 EST 1998


matrix proteins can be difficult to dissolve in sds buffers cause of
their large fibril structure, and for collagen, cause of their
triple helix.  solubility is often a problem.  a Tris buffer (pH 6.8)
with 4% SDS and 8M urea will be as good as you will get without alot
of sample preparation.  although phosphate buffers have superior
dissolving power than are often problematic with SDS due to insolubility 
of K PO4.

Jim Bassuk
U Washington
Seattle


On Tue, 24 Mar 1998, roberto wrote:

> We localized lectin binding sites on Xenopus laevis eggs. At the
> electron microscope such oligosacharides are asociated to the
> glycocalix. We just started biochemical experiments to study such
> glycoproteins by SDS-PAGE and Western-blotting with biotinylated lectins
> and streptavidin-peroxidase. The data suggest that some lectin reactive
> glycoproteins are lost during the sample preparation while others are
> retained. The preparation is suited for retaining plasma membrane
> proteins. By analizing pellets or supernatants which are intermediates
> in our purification method, we identified some protein bands reactive to
> the particular lectin we are interested.
> We're looking for a sample preparation suited for retaining both plasma
> membrane and the associated extracellular matrix or the latter only.
>             Thank You
> 
> 
> 
> 
> 




More information about the Proteins mailing list