Bio-Rad Comparative Proteomics Kit II: Western Blot Module - Manual (Page 5)

Lesson 3 Quick Guide
1. Using a ruler, chop the top and bottom off
the gel.
2. Equilibrate the gel in blotting buffer for
15 minutes on a rocking platform.
3. Soak fiber pads thoroughly in blotting buffer.
4. Mark the white nitrocellulose membrane
with penciled (or black ball point pen) initials
and prewet in blotting buffer along with the
blotting paper.
5. Make the blotting sandwich:
a. Add 1 cm depth of blotting buffer to
container and insert plastic cassette
with black side down.
b. Lay a wet fiber pad on the black side of
the cassette.
c. Lay one wet blotting paper on the fiber
pad and roll out air bubbles.
d. Lay gel squarely on blotting paper and
roll out air bubbles.
e. Lay wet nitrocellulose membrane on the
gel and roll out air bubbles.
f. Lay one wet blotting paper on the
membrane and roll out air bubbles.
g. Lay a wet fiber pad on top of the blotting
paper.
h. Close the cassette and clamp together
with the white clip.
29
Quick Guide
Q
UICK GUIDE
Fiber pad
Blotting paper
Membrane
Gel
Blotting paper
Fiber pad