I plan to run Mass Spectrometry on Tagged polypeptides using EasyPep kit from ThermoFisher for protein lysate prep!
I am wondering what is the best way to make a pellet of lysed worms. I see there are different methods such as sonication, metal and glass beads …but I man not sure which one is the most appropriate!
I appreciate to know about your experience or recommendation you might have. Please let me know!
We have used the Next Advance Bullet blender with 0.2mmstainless steel beads and setting of “8” for 3 mins to male lysates for Mass Spec. We have also used the Bioruptor Pico sonication system for making lysates. However not with the Easy prep kit.
I just realized now that you had answered my question. I used shock freezing in liquid N2, storage at -80, and then sonicated the pellet. It worked quite well and I got more protein out app 300 ul pellet than I could expect.