I am working with a parasite who total protein contents I have to analyse both by PAGE and Western blot. Can anyone suggest a protocol for going about this. I will be grateful. Thanks
I make protein extracts from Xenopus embryos using the embryos lysis buffer containing 150mM NaCl, 50mM Tris HCl pH7.5, 5mM EDTA, 5mM EGTA and 0.5%NP40. This works really well. You mesh the embryos using a pipette, vortex them hard and incubate on ice for 10min with intermittent vortexing. Post incubation, centrifuge them in a small, table-top, cold centrifuge at max speed for 10mins, and take the supernatant as your lysate.
This should work for you as well. In case your worms are a bit hard, you can add a bit of a stronger detergent and may be sonicate as well. But first I will try the protocol as it is.