Native protein extraction

[R. John Lye]

Aurelio, Oscar wrote:

> Can anyone point our group to a protocol to extract NATIVE protein 
> (i.e. no Tri-Reagent/Trizol phenol-solubilized protein) extraction 
> from C. elegans? We obviously wish to avoid using proteinase K to lyse 
> the worms. My colleague wishes to perform kinetic assays on an enzyme 
> from worms. Thanks very much in advance for any help.
>

I used a French press to homogenize the worms for enzyme assays.  I 
used a Pipes
based lysis buffer with lots of proteinase inhibitors - the worms are 
little bags of
proteinases!  More details in an ancient paper :

Lye, R.J., M.E. Porter, J.M. Scholey and J.R. McIntosh:  Identification 
and
Characterization of a Cytoplasmic Microtubule Translocator in the 
Nematode
C. elegans. Cell 51:309-318, (1987).

Hope that helps,
John