Vector Laboratories offers many enzyme substrate kits for use
with peroxidase, alkaline phosphatase, and glucose oxidase
detection systems.
All Vector Laboratories substrate kit reagents are supplied
in convenient dropper bottles promoting ease of handling
of chromogens and eliminating wait times for mixing and
dissolving powders or tablets.
Choose a substrate that matches the enzyme in your detection
system. Choosing the optimal substrate for your application
will depend on several considerations:
Sensitivity. Substrates provide varying degrees of sensitivities
(see sensitivity comparison chart below). For example,
ImmPACT™ peroxidase substrates are about 2-4 times more
sensitive than the original peroxidase substrate kits, and the
alkaline phosphatase substrate BCIP/NBT can achieve increased
sensitivities with longer incubation times.
Color. Color contrast is essential in multiple antigen labeling
applications, in pigmented tissues such as melanomas, and in
counterstained tissues. Color choices also depend on personal
preference.
Visualization. Viewing options include brightfield,
fluorescence, darkfield, electron microscopy, and spectral
imaging.
Heat resistance. For IHC/ISH double labeling applications, the
heat resistant substrate is applied first in IHC, followed by ISH
detection that includes a heat denaturation step.
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Multiple Antigen Labeling
Vector Laboratories is a leader in multiple antigen labeling
applications because of our wide range of very sensitive and
very low background detection reagents, our proprietary
enzyme substrates, and our rigorously tested protocols. These
detection systems and unique enzyme substrates offer an
extensive choice of color combinations for multiple labeling
using either:
The same enzyme system with different substrates to detect
each antigen, or
Different enzyme systems and their substrates to detect each
antigen.
The following Enzyme Substrate Combinations Table is designed as a reference for optimal multiple labeling, because the order of the two colored precipitates can significantly affect the quality, color, and labeling pattern of each antigen in the stained section. This chart ensures that distinct colors are visible after the labeling reactions are completed using an optimized multiple labeling protocol.
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