PBMCs were isolated by standard density centrifugation, by layering blood over an equal volume of lymphoprep (Axis Shields, Dundee, UK).
The 'buffy coat' layer was removed, re-suspended in RPMI media (Gibco, UK), and layered onto an equal volume of lymphoprep gradient solution (Robbins Scientific, UK).
Cells were then resuspended in Nycodenz-EDTA (Nycomed, Oslo, Norway) at 1.077 g/cm (5 ml per four mice) and layered onto an equal volume of fresh Nycodenz-EDTA of the same density, followed by an overlay of EDTA-FCS.
Blood was diluted 1 1 with RPMI and layered on an equal volume of Ficoll for density gradient centrifugation.
A layer-by-layer deposition of equal amounts of graphene oxide (GO) and the Ru II) complex on hydrophilic substrates through electrostatic attraction of these two components was evidenced by X-ray photoelectron and UV visible absorption spectroscopy.
Water-soluble and chloroform-butanol layer were separated by centrifugation, an equal amount of ethanol was added to the water-soluble layer (this procedure was repeated twice), and the ethanol-precipitated material was freeze-dried and stored at -80°C until use [ 19].
In native S-layers, carboxyl groups are neutralized by an equal number of amino groups, leading to a charge neutral outer surface (Weigert & Sára, 1995).
Moreover, native S-layer proteins from several Bacillus strains are comprised of carboxyl groups, which are neutralized by an equal number of amino groups and thus, leading to a charge neutral outer surface [29 31].
The lower PEMFA layer was removed and an equal volume of methanol was added to the remaining heptane solution.
