Whole protein extraction from tissues
Pre - cool lysis buffer on ice .
Add protease inhibitors just before using .
Add 10 fold lysis buffer to tissues ( 1ml lysis buffer / 100mg tissue ) .
Homogenize tissues with dispenser ( working time 10s / rest time 20s , repeat 5 times ) until no visible tissue blocks .
Mix samples on a rotating mixer at 4 °C for 2h .
Spin @ 16,000g , 10min , 4°C .
If necessary , centrifuge again .
Save supernatant .
Determine protein conc .
Make aliquots and store in - 80°C .
