Chemiluminescence and Bioluminescence, Analysis by
525
(f)
Alkaline phosphatase
OPO
3
Na
O
O
Cl
OMe
Cl
O
+
O
−
O
OMe
+
Light
Si
Si
Si
Si
Si
Si
H
O
H
O
H
O
H
O
O
O
H
H
H
2
O
2
Oxidation products
+
Light
(g)
N
N
H
Ph
Ph
Ph
O
2
/ HO
−
PhCON
=
CPh
−
NHCOPh
+
Light
(h)
N
N
Me
Me
+
+
H
2
O
2
Me
N
O
+
Light
(i)
Fig. 1
(
Continued
).
4
Applications
Most assays simply involve mixing the
sample with the appropriate CL or BL
reagents and measuring or detecting the
light emission. CL and BL reactions are
sensitive and simple to perform; they uti-
lize nonhazardous reagents, have a wide
dynamic range (several orders of magni-
tude), and are usable in diverse applica-
tions (assays for proteins, DNA, RNA, en-
zymes, substrates, inhibitors, metal ions,
and biomass) and in different assay for-
mats (e.g. flow injection, liquid chromatog-
raphy, extra-laboratory tests) (Table 2).
4.1
Measurement of Light Emission
Light can be measured quantitatively using
a luminometer or qualitatively by means
of photographic or X-ray Flm. Quantita-
tive measurements are usually performed
using a photomultiplier tube as the de-
tector. This device can be operated in
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