VectorCatalog2012.pdf? - Vector Laboratories

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V E C T O R L A B O R A T O R I E S
Enzyme Substrates
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.
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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.
Additional Information
• Ordering information on page A21.
• Substrate properties are summarized in the following
tables.
• Photomicrographs of each precipitated substrate and
additional descriptions can be found in Section I, “Enzyme
Substrates”.
• Counterstain compatibility of substrates can be found on
page A25.
• Substrate compatibility in multiple antigen labeling
applications can be found on page A22.
Relative Sensitivity of Substrates in Immunohistochemistry
HRP substrates were used with the ImmPRESS Polymer Detection Reagent. AP substrates were used with the VECTASTAIN ® ABC-AP Kit.
Peroxidase
(HRP)
Alkaline
Phosphatase
(AP)
SG
AEC
ImmPACT
SG
Competitor
DAB tablets
Vector ® Red
VIP
Vector ®
Blue
* Reaction product deposition not discreet and
can be variable (see full description in Section I,
“Enzyme Substrates” page I8)
** Longer incubation times increase sensitivity
DAB
(with Nickel)
DAB
ImmPACT
AEC
NovaRED
Sensitivity
BCIP/NBT
Vector ®
Black
ImmPACT
VIP
ImmPACT
AMEC Red
ImmPACT
DAB
ImmPACT
NovaRED
TMB
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*