Hippocampal and entorhinal cortex tissue samples were homogenized in 10 volumes of
RIPA lysis buffer (50 mM Tris, pH 7.5, 150 mM NaCl, 0.1 % sodium dodecyl sulfate and 0.5 % deoxycholate, and 1 % NP40) containing a cocktail of protease and phosphatase inhibitors (20 mg / ml each of pepstatin A, aprotinin, phosphoramidon, and leupeptin; 0.5 mM 4 -(2 - aminoethyl) benzenesulfonyl fluoride hydrochloride; 1 mM EGTA; 5 mM fenvalerate; and 5 mM cantharidin).
Lane 1: HEK cells Lane 2: hfRPE cells Lane 3: hfRetina Lane 4: Mouse liver Lane 5: Mouse RPE Lane 6: Mouse retina Lane 7: Zebrafish liver Lane 8: Zebrafish retina PVDF membrane
RIPA lysis buffer with mixed phosphatase and protease inhibitor cocktail