Which are the best cells breakage methods in small-scale expression screening of recombinant protein in P. pastoris? I need to lyse the cells to perform western blot analysis.
I tried with glass beads, but with this small volume (2 ml culture) it looks like the cells are still whole.
Do you think I can boil the collected cells directly in SDS-page sample buffer?
León Samaniego
Hola Valentina. With chemical methods: Zymolyase (digests cell walls) and lysis buffer (SDS) both give a good lysate. Physical method can use, glass beads with vortex or also sonication.
And why not use a secretion system in P. pastoris?
0 votes
0 thanks
- Badges
- Science method
Similar questions and discussions