3
6
D. V. Yanvarev et al.
suspended in 5 mM Tris-HCl buffer (200 µl, pH 7.5). The cells were
degraded by the fourfold freezing-melting procedure in liquid nitrogen,
methanol (400 µl) was added, and the resulting suspension was centrifuged
(
2 minutes at 14,000 g). The supernatant was concentrated in vacuum
◦
at 37 C. The residue was dissolved in TEAB (25 µl) and analyzed by
HPLC.
The lipid composition of the membrane fraction was analyzed by silica
gel TLC with unlabelled authentic samples of the corresponding lipid
components as reference compounds. Phospholipids were detected by the
Vaskovsky and Kostevsky method.[
13]
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