Bio-Rad Immun-Star™ AP Chemiluminescence Kits - Manual (Page 21)

9. Blot development – remove the membrane and drain
excess liquid without letting the blot dry. Place on a piece of
Saran wrap on a level surface. Pipet the substrate
solution onto the membrane. Incubate 5 min. Remove the
membrane, draining the excess liquid from the blot. Seal in
a heat-sealable bag or in Saran wrap. Do not allow the
membrane to dry, because this will result in a loss of signal.
10. Film exposure/imaging – expose X-ray or instant film
to the blot at RT. Exposure time will depend on the type of
membrane used and the protein concentration. Table 1
lists recommended exposure times; however, optimal
conditions should be determined for each particular appli-
cation. Develop the film according to the manufacturer’s
instructions. Alternatively, a Bio-Rad imager such as the
Bio-Rad VersaDoc
™ or ChemiDoc ™ system can be used
to capture the signal.
Table 1. Typical exposure times for nitrocellulose
and PVDF membranes.
With Immun-Star Without
Membrane Enhancer Enhancer
Nitrocellulose 1–10 min 12–24 hr
PVDF Not recommended 1–10 min
18