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Brain samples were thawed from -80°C storage to -8°C in cryostat.
Brain samples were thawed and homogenized in ice to ensure the viability of proteins using a Duall, all-glass, tissue grinder.
Complete equilibration was observed by Day 1 and no degradation in quinacrine was noted through day 7. Brain samples were thawed (in plastic tubes) at room temperature for 15 20 min and weighed.
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Frozen brain tissue samples were thawed, washed with ice-cold phosphate-buffered saline (PBS), suspended in radio-immunoprecipitation assay (RIPA) buffer containing protease inhibitors (2 μg/ml of aprotinin, 10 μM of leupeptin, 1 mM of phenylmethylsulfonyl fluoride), and minced on ice.
Snow samples were thawed overnight at 21 °C.
EV samples were thawed in 37 °C water immediately prior to analysis and diluted (50 100×) in DPBS.
To determine the best thawing method, handling regimen 1 through 7 samples were thawed using two methods.
The plasma samples were thawed at room temperature.
Frozen tissue samples were thawed for 45 min and homogenized.
The frozen tissues from the initial samples were thawed.
For NMR analysis, serum samples were thawed on ice.
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