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Featuring:
Plus
quality reagents for:
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| DuoLuX Chemiluminescent and Fluorescent Peroxidase Substrate
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The DuoLuX™ Chemiluminescent/Chemifluorescent Substrate
is a unique formula based on acridan chemistry. The DuoLuX™
Substrate provides the following advantages:
Very high sensitivity
Prolonged and intense light emission
Chemiluminescent or fluorescent detection
Permanent fluorescence
The DuoLuX™ Chemiluminescent/Chemifluorescent Substrate
is supplied for either horseradish peroxidase or alkaline
phosphatase development and can be used for western,
Southern, northern, or dot blots, or for ELISA applications. This
substrate is about 10 times more sensitive than the alkaline
phosphatase substrate BCIP/NBT and about 100 times more
sensitive than the peroxidase substrate DAB.
The DuoLuX™ Chemiluminescent/Chemifluorescent
Substrate for peroxidase (SK-6604) is supplied in two
bottles, consisting of 100 ml of the DuoLuX™ Substrate
(Reagent 1) and 100 ml of a peroxidase converter solution
(Reagent 2). Reagents 1 and 2 are mixed in equal volumes just
prior to use, providing 200 ml of peroxidase substrate working
solution.
Chemiluminescent properties
Light emission of the DuoLuX™ Chemiluminescent/
Chemifluorescent Substrate occurs continuously over a
long period of time for both the peroxidase and alkaline
phosphatase substrates. With either substrate formula,
reacted DuoLuX™ Substrate luminesces in the blue range with
a peak emission at 453 nm. Unlike other chemiluminescent
substrates, blots can be reexposed to film many times over an
8 hour period to optimize band intensities or resolution.
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Serial dilutions (1:2) of mouse IgG were resolved by PAGE, transferred onto nitrocellulose membrane and detected with the DuoLuX™ Substrate using either the Alkaline Phosphatase-based VECTASTAIN® ABC-AmP™ Kit (lower left) or Peroxidase Streptavidin (lower right). Graph shows prolonged light emission characteristics of the DuoLuXTM Substrate with each enzyme.
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Fluorescent Properties
In addition to its chemiluminescent properties, the reaction
product of the DuoLuX™ Substrate is also fluorescent.
Fluorescence can be recorded with a digital imaging system
or a conventional camera months after chemiluminescence
has faded. The excitation maximum is at 405 nm, but other
wavelengths (254 nm and 365 nm) also excite. Maximum
fluorescent emission occurs at 453 nm. For fluorescence
detection, nitrocellulose is recommended due to the
intrinsic fluorescence of nylon. Acquisition of fluorescent
signal requires a much shorter exposure time than
chemiluminescence, often a fraction of a second.
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Western blot visualized by fluorescence using the VECTASTAIN® ABCAmP™ Kit with the DuoLuX™ Substrate. Serial dilutions (1:2) of rabbit
IgG heavy chain were resolved by PAGE, transferred onto nitrocellulose
membrane, and detected with biotinylated anti-rabbit IgG, followed
by the VECTASTAIN® ABC-AmP™ Kit with the DuoLuX™ Substrate. [A]
Fluorescent detection by image acquisition (1 second). [B] Fluorescent
image of the same blot as shown in [A] obtained 12 months later.
(Same 1 second acquisition time.)
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Additional Information:
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Application: Nucleic Acid Detection
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Application: Protein Detection
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