Nupage 4 12 bis tris zoom gel

The comparison between hand-cast and pre-cast gels, as well as different SDS-PAGE gel percentages and sizes were also made in this study. The conventional protocol involved hand-casting gradient gel of 8–15%. Glass plates of 16 x 18 cm, with a 1 mm thick gradient gel was prepared using a gradient maker (Model SG 30, Hoefer, USA). Agarose sealing solution of 0.5% was used and electrophoresis was performed by SE 600 Ruby Electrophoresis System (GE Healthcare, Uppsala, Sweden) linked to a cooling circulator (Grant Instrument Ltd., Cambridge, UK) and Power Supply-EPS601 (GE Healthcare) at 18°C. The gel was run using SDS electrophoresis buffer (25 mM Tris; 198 mM glycine; 0.1% SDS) under these conditions: (i) Phase 1: 50 V, 150 mA, 100 W for one hour; (ii) Phase 2: 600 V, 150 mA, 100 W until the tracker dye reached the bottom of the gel. In the modified protocol, pre-cast NuPAGE 4–12% Bis-Tris ZOOM Gel, size of 8 x 8 cm (Invitrogen, California, USA) was used and electrophoresis was performed in the Mini-PROTEAN Tetra Vertical Electrophoresis Cell (Bio-Rad). Similarly, 0.5% agarose sealing solution was used, but NuPAGE MOPS SDS Running Buffer (1X) was used to run the electrophoresis at 200 V, 2 mA instead. The electrophoresis was stopped when the marker reached the mark above the protrusion of the gel cassette.