CHAPTER 3
18
Operation
It is very important that the glass and silicone mat be free of smudges,
dust, and dye before placing membranes or gels onto the Odyssey
®
Imager. Contaminated surfaces in contact with the membrane
surface may cause blotches and streaks that cannot be removed with
further washing.
For Western blotting methods, nitrocellulose or PVDF membranes
may be used (see
www.licor.com
for the latest membranes and kits).
For best performance, pure cast nitrocellulose membranes are
recommended. Detailed blotting protocols can be found in the
Odyssey Application Protocols. There are some general tips,
however, for using membranes with the Odyssey Imager.
•
Do not touch the membrane – handle only with a clean, smooth-edged
forceps. Lift the membrane only by the corners. Fingerprints, even from a
glove, will show clearly when imaged.
•
Use the silicone mat included to cover the membrane(s) before scanning.
Use the 4" soft roller included to remove any air bubbles that may be
present. These optional steps help keep the membrane flat against the
scanning surface for optimum imaging.
•
Protect the membrane from light until it has been scanned.
•
Keep the membrane wet if it is to be stripped and re-used. For Western
blots, store dry or in PBS buffer at 4°C.
•
Use clean containers to avoid cross-contamination and reduce
background.
•
Multiple membranes can be washed together, provided there is ample
volume so each membrane moves freely.
•
If the signal on the membrane is too strong, re-scan at a lower intensity
setting.
•
The fluorescent signal on the membrane will remain stable for several
months or longer if protected from light.
Using Membranes
