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A context test was performed in the same conditioning chamber for three min in the absence of CS.
After stirring, the acidic solution was centrifuged at 9000 g for three min to remove the precipitate (mainly mucins) and the acidic solution was immediately analyzed by HPLC-ESI-MS (100 µL, corresponding to 50 µL of whole saliva) or stored at −80°C until the analysis.
The column was then kept at 100% methanol for three min. The column was reconditioned in 25% methanol for two min. The electrospray interface was operating in the positive ion mode at 370°C and the ion spray voltage was −4000 volts.
For FOXO3a staining, HOZOT cells were washed in PBS and cytospun onto glass slides in a Cytospin 2 (SHANDON Inc., Pittsburgh, PA) at 700 rpm on high acceleration for three min. Cytospins were fixed in 4% paraformaldehyde for 20 min at room temperature.
Each solution was sequentially infused for three min with no pauses between the infusions.
16S rRNA FISH - Biopsies were incubated for three min at room temperature in 50%, 80% and 100% ethanol, respectively.
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£15 for one rider, £26 for two, min. height 1.2 metres, rockreef.co.uk Castle Keep and Black Gate reopen on 21 March as Newcastle Castle, after a £1.67m refurbishment.
When the first number-symbols were written down some 5,000 years ago in Sumer, a region of what is now Iraq, the word for one (ges) also meant man, or erect phallus, while the word for two (min), also meant woman.
The reduction in absorbance was measured spectrophotometrically after one minute of mixing the solutions and continued for six min. Ascorbic acid was used as a positive control.
Aerosol exposures were conducted for ten min, followed by 5 min purging with air.
The cells were fixed in 4% paraformaldyhyde for 15 min followed by three washes with PBS for five min each.
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