The protein was then desalted into spectroscopy buffer using a Disposable PD-10 Desalting Columns (GE Healthcare Life Sciences, UK).
The process was repeated once again, and the protein solution was then desalted into the rehydration buffer (8 M urea, 2% CHAPS, 0.5% IPG buffer, pH 3−10) by the Millipore Centricon YM-10 as salt-free 2-DE protein samples.
An aliquot (70 μL) was then desalted into ammonium acetate buffer (20 m m, pH 7.0), and analysed by ESI-MS to ensure complete conversion of K165C into K165Dha.
rOmpF was then desalted with 20 mM Tris HCl buffer [pH 7.4, containing 150 mM NaCl and 0.1% (v/v) LDAO] using a HiPrep 26/10 desalting column.
The desired protein was then desalted and kept in the storage buffer (25 mM Tris-HCl, pH 8).
The crudely purified sample was then desalted in 20 mM Bis-Tris pH 6.5 buffer using two HiPrep 26/10 Desalting columns in series before loading onto a Tricorn 10/100 anion exchange column packed with Source 15Q resin.
The media containing the protein was then desalted using a HiPrep 26/10 desalting column (GE Healthcare Life Sciences) equilibrated with Buffer A (20 mM phosphate, pH 7.4).
The media containing the protein was then desalted using a HiPrep 26/10 desalting column (GE Healthcare Life Sciences) equilibrated with Buffer A (20 mM Tris-HCl, pH 8.5).
The POSWOI-rich fraction was then desalted using HiTrap desalting column (GE Healthcare) and changed to 0.05 mol/L sodium acetate buffer (pH 4.8).
100 μl of the boiled extract was then desalted using a 0.5 ml Zeba column (Pierce; pre-equilibrated with 3 washes of Tris-EDTA buffer, pH 8), using a syringe to displace the DNA.
