In order to remove imidazole and NaCl from the eluate, the pooled elution fractions were loaded to a 60 mL size exclusion column (Sephadex G25 matrix), which was equilibrated with sodium phosphate buffer (50 mM, pH 7.5) before.
The agarose binding assay using high molecular mass (HMM) and low molecular mass (LMM) AID was performed as described above; instead of unfractionated cell lysates, 100 µl of the relevant 1 ml size exclusion chromatography fractions were used.
The 9 ml size lysis buffer tube was used.
One milliliter of the solution was transferred into an Eppendorf tube (1.5 mL size).
Machines typically included sports drinks, flavoured, sweetened waters and iced teas in 500 ml to 600 ml sizes.
Local compliances were analyzed as the local slope (dV/dp) within consecutive volume windows of 50 ml size.
Sterile water was added and the formulated mixture was filtered through a sterile Millipore filter (0.22 μm, 4 mm) directly into a sterile product vial (10 mL size).
Chambers of 20 mL size containing Krebs-Ringer buffer of the following composition (mM) were used: 119 NaCl, 25 NaHCO3, 11.1 glucose, 1.6 CaCl2 × H2O, 4.7 KCl, 1.2 KH2PO4, 1.2 MgSO4 × 7 H2O.
Separation of cluster compounds were conducted on an ÄKTApurifier HPLC system equipped with a Superdex 75 PC 3.2/30 (2.4 mL) size exclusion column.
Then, the algae were washed off from the steel mesh and evenly distributed among 15 to 25 Erlenmeyer flasks (150 ml size) filled with ~40 ml Volvox liquid medium.
