Serum samples were separated on 4%–15% Tris-Glycine precasted gels (BioRad), for the denaturant PAGE and on NativePAGE Bis-Tris Gel System 4%–16% (Invitrogen, Thermo Fisher Scientific), for the native PAGE, according to the manufacturer's instructions.
In both cases, serum proteins were transferred from the gel to PVDF membranes and probed with anti-human ApoA-I antibodies (64308, Abcam, for denaturant blot and Q0496, Dako, Agilent Technologies, for native blot). Detection was performed by using HRP-conjugated secondary antibodies (GE Healthcare) and a chemiluminescence detection substrate (Super-Signal West Femto, Thermo Fisher Scientific). Blots were imaged using the Odyssey Fc system (LI-COR Biosciences).
In both cases, serum proteins were transferred from the gel to PVDF membranes and probed with anti-human ApoA-I antibodies (64308, Abcam, for denaturant blot and Q0496, Dako, Agilent Technologies, for native blot). Detection was performed by using HRP-conjugated secondary antibodies (GE Healthcare) and a chemiluminescence detection substrate (Super-Signal West Femto, Thermo Fisher Scientific). Blots were imaged using the Odyssey Fc system (LI-COR Biosciences).