Brains were immediately removed and tissues were dissected on ice. Tissues were frozen on dry ice and kept at -80 °C until they were homogenized. The cortices were homogenized without thawing in 5 times volume-weight of RIPA with a mechanical homogenizer (14-261-01, Fisher Scientific, Waltham, MA, USA). The hippocampi were homogenized without thawing by sonication in 150µL of RIPA. Samples were then centrifuged for 20 min at 20,000g at 4 °C. The supernatant was recovered, diluted in sample buffer (NuPAGE LDS; Invitrogen, Carlsbad, CA) containing 5% of 2-β-mercapto-ethanol, 1 mM Na 3 VO 4 , 1 mM NaF, 1 mM PMSF, 10 μl/ml of Proteases Inhibitors Cocktail (P8340; Sigma-Aldrich, St. Louis, MO, USA), and heated for 10 min at 95 °C. 10 μg-20µg of protein were analyzed as described previously (Petry, Pelletier et al. 2014 , Petry, Nicholls et al. 2017) .
Mechanical homogenizer
Manufactured by Thermo Fisher Scientific
Sourced in United States
A mechanical homogenizer is a laboratory instrument used to blend and mix samples or solutions into a homogeneous mixture. It typically consists of a motorized device with a rotating shaft and a blade or rotor that disrupts and disperses solid or semi-solid materials within a liquid.
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Lab products found in correlation
2 protocols using mechanical homogenizer
1
Protein Extraction from Brain Tissues
2
Protein Extraction from Brain Tissues
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Brains were immediately removed and tissues were dissected on ice. Tissues were frozen on dry ice and kept at -80 °C until they were homogenized. The cortices were homogenized without thawing in 5 times volume-weight of RIPA with a mechanical homogenizer (14-261-01, Fisher Scientific, Waltham, MA, USA). The hippocampi were homogenized without thawing by sonication in 150µL of RIPA. Samples were then centrifuged for 20 min at 20,000g at 4 °C. The supernatant was recovered, diluted in sample buffer (NuPAGE LDS; Invitrogen, Carlsbad, CA) containing 5% of 2-β-mercapto-ethanol, 1 mM Na 3 VO 4 , 1 mM NaF, 1 mM PMSF, 10 μl/ml of Proteases Inhibitors Cocktail (P8340; Sigma-Aldrich, St. Louis, MO, USA), and heated for 10 min at 95 °C. 10 μg-20µg of protein were analyzed as described previously (Petry, Pelletier et al. 2014 , Petry, Nicholls et al. 2017) .
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