Freezing worms for western blots

Model Organism: Worms Methods & Reagents
1

I would like to freeze my worms for western blots at a later time. Wondering what my options are?

  1. Should I be freezing them in the Laemmli buffer (before or after boiling?)
  2. Should I freeze them in RIPA (a lysis buffer) and then add the Laemmli Sample buffer after the thaw?

Any help appreciated!

2

I have had the best success with processing the worms all the way (i.e., through boiling in Laemmli buffer) and then immediately freezing. This insures that any proteases are degraded before freezing. With this method, I have successfully found even non-abundant proteins with a Western protocol after months at -30C.

Janet