HELP: protocol for protein extraction - (Mar/11/2008 )

I have to extract proteins from yeast, and then run a Native PAGE gel. If anyone have that protocol for protein extraction?
Thanks in advance!

I am not sure about yeast but for mammalian cells, we use the following triton buffer to make extracts.

TBS-Triton extraction buffer
50 mM Tris ph 7.5
150 mM NaCl
0.1% Triton X-100
10mM NaF
Add DTT to 1.5mM and PI to 1X immediately before extraction

The cells are washed before the buffer is added

Cells are sonicated and centrifuged

Supernanant and pellet are saved.

0.1% Triton X-100
Add DTT to 1.5mM and PI to 1X immediately before extraction

If these reagent would destroy structure of proteins?

These should be fine as we use this buffer for some protein assays. the protein activity is preserved. We have done this for many different enzymes preparations.

Thanks! I'll have a try.