Livers were homogenized in extraction buffer (20 mM Tris-HCl, 250 mM saccharose, pH 7.2, 2 mM EDTA, 40 mM KCl and Roche Complete Protease Inhibitor Cocktail) using an Ultraturax homogenizer.
The tissue was then spun down for 5 min at 1,000 × g and briefly homogenized in ice-cold PBS supplemented with protease inhibitor cocktail using an Eppendorf-fitting pestle homogenizer.
To study the impact of the microbiota on spontaneous contractile behaviour, we measured the contraction frequency in germ-free (GF) animals which were obtained by treating H. vulgaris polyps with an antibiotic cocktail, using a regime known to completely remove associated bacteria52.
As they did this, Matthew allegedly fashioned a molotov cocktail using a curtain and a jar of home made moonshine they had with them.
As they did this, Matthew allegedly fashioned a molotov cocktail using a curtain and a jar of homemade moonshine they had with them.
The system was calibrated with O2− obtained from the UV photolysis of Benzophenon and 2-Propanol pH 12 cocktail using a mercury pan-lamp (Pen Ray; [27]; Fig. S1).
350 μL of the supernatant of this second spin was counted in 5 mL of scintillation cocktail using a scintillation counter.
BV-2 cell pellets (1×10 cells) were homogenized in 1 ml STM solution (0.25 M sucrose, 10 mM Tris HCl, 1 mM MgCl2, and 1X protease inhibitor cocktail) using a syringe with a 261/2-gauge 261/2-gauge
BV-2 cell pellets (1×10 cells) were homogenized in 1 ml high-salt buffer (2 M NaCl, 10 mM HEPES-NaOH, pH 7.4, 1 mM EDTA, and 1X protease inhibitor cocktail) using a syringe with a 261/2-gauge 261/2-gauge
To analyze the ubiquitination of GluA1, samples of barrel cortex were collected and homogenized in lysis buffer (150 mM NaCl, 20 mM HEPES, 4 mM EDTA, 1 mM EGTA, and 1% Triton X-100, with a protease inhibitor tablet and phosphatase inhibitor cocktail) using a tissue homogenizer.
To prepare the crude lysates, frozen samples were homogenized in ice-cold homogenization buffer B (150 mM NaCl, 20 mM HEPES, 4 mM EDTA, 1 mM EGTA, and 1% Triton X-100, with a protease inhibitor tablet and phosphatase inhibitor cocktail) using a glass/glass tissue homogenizer.
