Bio-Rad Immun-Star™ AP Chemiluminescence Kits - Manual (Page 5)

of the antigen in a polyacrylamide or agarose gel, passively by
directly spotting the antigen onto a membrane, or by vacuum
filtration using a microfiltration apparatus. Following antigen
binding, the remaining protein binding sites on the membrane
surface are blocked with nonfat dry milk, or other protein
blocking agents.
The membrane with bound antigen is then incubated with a
primary antibody specific for the antigen to be detected. The
blot is washed to remove unbound antibody and incubated
with the respective second antibody, which has been
conjugated to alkaline phosphatase (AP). The membrane is
then treated with the chemiluminescent substrate alone for
PVDF membranes, and with substrate and enhancer for
nitrocellulose. The concentrated enhancer is added to the 1x
substrate solution and does not add any extra incubation or
wash steps. The blot is then used to expose X-ray or instant
film, or imaged by an imager capable of detecting chemilumines-
cent signals, such as the Bio-Rad VersaDoc
™ or ChemiDoc ™
system.
1.3 Immun-Star Products
Catalog # Description
170-5010 Immun-Star Goat Anti-Mouse -AP Detection Kit
170-5011 Immun-Star Goat Anti-Rabbit -AP Detection Kit
170-5012 Immun-Star AP Substrate Pack
170-5018 Immun-Star AP Substrate
170-5013 Immun-Star Goat Anti-Mouse -AP Intro Kit
170-5014 Immun-Star Goat Anti-Rabbit -AP Intro Kit
2