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The study itself is fairly straightforward and includes a gradual, measured ascent to altitude and a series of tests undertaken prior, during and after ascent and the descent following the expedition.
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And it measured the ascent to the Presidency of a brute with authoritarian tendencies.
After 30 min vibration, with 1× substrate and stop buffer each prepared, luciferase activity was measured by Ascent software on chemiluminescent plate reader (Thermo).
Labelled cells were detached (Trypsin-EDTA) from the underside of ThinCerts and fluorescence measured (Fluoroskan Ascent FL plate reader, Cheshire, UK).
Optical densities (405 nm and 540 nm) were measured (Multiskan Ascent plate reader, Cheshire, UK) and the absorbance of cells without BrdU subtracted from each reading.
Luciferase activity was measured using Fluoroskan Ascent Luminometer (Lab Systems).
Luminescence was measured using Luminoskan Ascent (Thermo Scientific).
The fluorescence intensity was measured by Fluoroskan Ascent FL (Thermo Electron Corporation, Waltham, MA, USA) with the setting of 485 nm (excitation) and 530 nm (emission).
100 μl diluted supernatant and DCFH-DA (10 μM) were mixed and incubated at 37 °C for 30 min, Then, the fluorescence intensity was measured by Fluoroskan Ascent FL.
The caspase activity was luminometrically measured by Luminoskan Ascent microplate reader (Thermo Scientific, Vienna, VA, USA) and readings were normalized by total protein.
In detail, DNA from each placenta was measured with Fluoroskan Ascent (Thermo Labsystems, Franklin, MA) and Picogreen reagents and kits (Molecular Probes, P-7589).
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