Chemiluminescence and Bioluminescence, Analysis by
529
N
CO
O
Me
+
FSO
3
−
Labeled probe
N
CO
O
Me
+
FSO
3
−
Hydrolysis
No light
No hydrolysis
(protected)
Light
Target DNA
Borate buffer
Peroxide
+
NaOH
Fig. 4
Hybridization protection assay for detecting target
nucleic acid (DNA) using a chemiluminescent acridinium
ester label.
O
O
OMe
OPO
3
Na
2
R
CO
NH
NH
CO
NH
2
+
OH
I
+
H
2
O
2
Luminol
4-Iodophenol
DNA: DNA-biotin: SAV–AP
Membrane
AMPPD, R
=
H
CSPD, R
=
C
Light
Membrane
Light
DNA: DNA-HRP
(a)
(b)
Fig. 5
Chemiluminescent detection of an alkaline phosphatase label using an
adamantyl 1,2-dioxetane aryl phosphate (AMPPD
,CSPD
), and detection of
a peroxidase label using the enhanced chemiluminescent reaction. AP, alkaline
phosphatase; HRP, horseradish peroxidase; SAV, streptavidin.
blot,
Southern
blot),
the
nature
of
t
h
em
em
b
r
a
n
ei
n
fl
u
e
n
c
e
st
h
eC
Ls
i
g
-
nal from bound alkaline phosphatase.
The surface of a nylon membrane acts
as an enhancer of the CL signal be-
cause a dephosphorylated intermediate
is sequestered in hydrophobic domains
on
the
nylon.
Similar
improvements
