Peroxidases are a group of heme-containing enzymes that catalyze
oxidation-reduction reactions. Hydrogen peroxide (H2O2)
or lipid peroxides are the substrates used by peroxidases for a number of
oxidative reactions. Peroxidases are found in bacteria, fungi, plants and
animals. Cellular peroxidases play an important role in protecting cell from
oxidative injury. Horseradish peroxidase (HRP) is extensively used in ELISA
and IHC as a reporter molecule when conjugated with secondary detection reagents.
AnaSpec is pleased to introduce two luminescent
read-out assay kits for determining peroxidase activity – the SensoLyteŇ
Luminescent Peroxidase Assay Kit and the SensoLyteŇ Luminescent Peroxidase ELISA Assay
Kit. The former detects peroxidase activity in biological
samples and in ELISA with HRP-conjugates while the latter detects HRP conjugated with antibody or streptavidin
in ELISA. In both assays, the chemiluminescent substrate luminol is oxidized
by peroxide and creates an excited state intermediate product that decays
to a lower energy state by releasing photons of light.
The kits
provide ample materials to perform 500 assays in a 96-well format. The protocol
can readily be modified to run assays in a 384-well format.
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Figure 1. Detection of horseradish peroxidase with the SensoLyteŇ Luminescent Peroxidase
Assay Kit. The detection limit can reach femtogram range (n=3, mean±S.D).
Figure 2. The SensoLyteŇ Luminescent Peroxidase
ELISA Assay Kit was used to detect goat anti-rabbit biotinylated antibody.
Wells were coated with total rabbit IgG. After blocking and washing, serially
diluted goat anti-rabbit antibody conjugated with biotin was added into the
wells, followed by incubation with HRP-labeled streptavidin. The plate was
washed again and the peroxidase substrate mixture added to wells. Signal was
immediately measured (Flexstation 384II, Molecular Devices). The assay can
detect as low as 1 pg of biotinylated goat-anti-rabbit antibody.
Related Products
SensoLyte®
ADHP Hydrogen Peroxide Assay Kit *Fluorimetric*
SensoLyte®
ADHP Peroxidase Assay Kit *Fluorimetric*
