Bio-Rad Trans-Blot® Cell - Manual (Page 7)

Section 2
Trans-Blot Cell Assembly
2.1 Preparation for Blotting
1. Prepare the transfer buffer. (See Section 3.3 for buffer formulation. Using buffer chilled to
4 °C will improve heat dissipation.)
2. Cut the membrane and the filter paper to the dimensions of the gel. Always wear gloves
when handling membranes to prevent contamination. Equilibrate the gel and soak the
membrane, filter paper, and fiber pads in transfer buffer (15 min—1 hour depending on
gel thickness).
3. Prepare the gel sandwich.
Place the cassette, with the gray side down, on a clean surface.
Place one pre-wetted fiber pad on the gray side of the cassette.
Place a sheet of filter paper on the fiber pad.
Place the equilibrated gel on the filter paper.*
Place the pre-wetted membrane on the gel.*
Complete the sandwich by adding a piece of filter paper on the membrane.*
* Removing any air bubbles which may have formed is very important for good
results. Use a glass tube to gently roll air bubbles out.
Add the last fiber pad.
4
Fiber pad
Filter paper
Membrane
Gel
Filter paper
Fiber pad
Cathode (-) side
Anode (+) side