Trans blot turbo transfer cell

Proteins were electroblotted onto a low-fluorescent PVDF membrane using a Trans-Blot Turbo transfer cell and the RTA transfer kit (Bio-Rad Laboratories) for 10 min (25 V, 1.3 A, RT) and processed as described [31 (link)]. For detection, the membrane was incubated with Immuno Blue HRP-Substrate (NH DyeAGNOSTICS, Halle, Germany) for 10 min or with WesternBright Sirius HRP-Substrate (Advansta, Menlo Park, CA, USA) for 2 min, washed in Advan washing solution, and the fluorescence or chemoluminescence recorded (ChemiDoc MP CCD camera system, Bio-Rad Laboratories).

Proteins were electroblotted onto a PVDF membrane using a Trans-Blot Turbo transfer cell (Bio-Rad Laboratories) for 10 min (25 V, 1.3 A, RT). Membranes were blocked with Advan Blocking solution (30 min, RT) and incubated with serum samples (3 μL serum in 10 mL Advan Blocking solution) overnight at 4 °C. Afterwards, membranes were washed two times with Advan Washing solution (5 min), incubated with peroxidase-conjugated goat anti-mouse IgG + IgM (0.5 μL in 10 mL Advan blocking solution plus 0.5% BSA (w/v), 90 min, RT), and washed again with Advan Washing solution (3×, 5 min each). The membrane was incubated with Immuno Blue HRP-Substrate (NH Dyeagnostics, Halle, Germany) for 10 min, washed in Advan washing solution and the fluorescence was recorded (ChemiDoc MP CCD camera system, Bio-Rad Laboratories).