Oxidases,
present in many cells, are used in a wide variety of bioassays. The most useful known oxidase is horseradish
peroxidase (HRP), which has been coupled with sensitive fluorogenic and
chromogenic substrates to quantitate intracellular hydrogen peroxide production
as well as a diverse assortment of analytes including glucose, galactose,
cholesterol, and uric acid. Fluorogenic
peroxidase substrates are converted to highly fluorescent products in the
presence of the enzyme and hydrogen peroxide but can be relatively unstable for
use in enzyme-linked immunosorbent assays (ELISAs). By forming the diacetate derivates, (see M0807) intracellular applications are more
efficient, whereupon the acetates are cleaved by endogenous esterases,
releasing the intact substrate.1-5
FLUOROGENIC
SUBSTRATES
|
PRODUCT
# |
PRODUCT
NAME |
COLOR OF
RELEASED PRODUCT (Abs/Em) (nm) |
|
Dihydrorhodamine 123 |
Yellow-Green (505/534 (MeOH)) |
|
|
2’,7’-Dichlorofluorescin diacetate |
Green (495/529) |
|
|
Live Cell Fluorescent Reactive Oxygen Species Detection Kit (available soon!) |
Green (495/529) |
CHROMOGENIC
SUBSTRATES
|
Methyl Purple, Sodium Salt |
N/A |
REFERENCES:
1.
Anal Biochem 11, 6 (1965)
2.
J Clin Invest 87, 711 (1991)
3.
J Lab Clin Med 117, 291 (1991)
4.
Anal Biochem 187, 129 (1990)
5.
Anal Biochem 134, 111 (1983)