Cells were lysed with TN1 lysis buffer (50 mM Tris, 125 mM NaCl, 1 % Triton X-100, 10 mM EDTA, 10
mM sodium fluoride and 10 mM sodium pyrophosphate) supplemented with protease inhibitor cocktail (1.2 mM AEBSF, 0.46 μM aprotinin, 14 μM bestatin, 12.3 μM E-64, 112 μM leupeptin, 1.16 μM pepstatin)(Amresco; Solon, OH)-RRB- and 1 mM sodium orthovanadate (Enzo Life Sciences; Farmingdale, NY).
Not exact matches
Briefly, retinal samples were homogenized at 4 °C in buffer containing 10
mM Tris (pH 7.4), 1
mM EDTA, 0.15 M
sodium chloride, 0.5 % NP - 40, with protease inhibitors (20 µg / ml aprotinin, 5 µg / ml leupeptin, and 1
mM phenylmethane sulfonyl
fluoride - PMSF).
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).
Cells were lysed 48 h later with modified radioimmunoprecipitation assay buffer (RIPA buffer, 50
mM Tris - HCl, pH7.4, 150
mM NaCl, 1 % Nonidet P - 40 (NP - 40), 0.25 %
sodium deoxycholic acid, 1
mM ethylenediaminetetraacetic acid (EDTA), 1
mM phenylmethylsulphonyl
fluoride (PMSF), 1X Complete cocktail protease inhibitor (Roche, Penzberg, Germany)-RRB-.