Alkaline Phosphatase (AP) Substrates

Vector offers alkaline phosphatase (AP) substrates in both original and ImmPACT^® formulations.

The original Vector^®AP substrates offer a range of color choices and provide the following:

• Allow you to stain antigens in pigmented tissues and melanomas
• Enable localization of multiple antigens in a single tissue section (the combination of Vector^® Red and Vector^® Blue substrate, for example, are useful in co-localization experiments)
• Vector^® Red and Vector^® Blue substrates can be viewed with both conventional light microscopy as well as fluorescence microscopy
• Vector^® AP substrates can be both non-aqueous and aqueously mounted with the exception of Vector^® Black, which should only be non-aqueously mounted
• Vector^® Blue substrate generates a product that can be cleared with citrus oil-based clearing agents and then permanently mounted
• The BCIP/NBT substrate is frequently used for nitrocellulose or in situ hybridization applications

ImmPACT^® Vector^® Red Substrate is the most sensitive red AP substrate:

• 2-4 fold greater sensitivity than the previous version
• Supplied with stock solutions in dropper bottles and optimized diluent
• Optimized specifically for immunohistochemistry applications
• Can be viewed with fluorescence as well as light microscopy

Enzyme Substrates / Microscopy Table

This table is designed as a reference to help you determine the optimal mounting medium for your application.

Additional Enzyme Substrate Characteristics

This table is designed as a reference to help you determine the optimal mounting medium for your application.

Resources and Guides:

IHC Multiplexing Guide

A valuable resource for those considering localizing two or more antigens in the same tissue section. Protocols for chromogenic detection are accompanied by schematic illustrations of the procedures.
A chart of all pair-wise combinations of substrates is included along with photographic examples. Practical tips regarding order of substrates, blocking, and proper controls are also provided.

Enzyme Substrates Brochure

In addition to photomicrographs that help the user visualize the color that is generally produced
by each substrate, this brochure contains information on substrate properties that are especially useful for those who wish to multiplex (or introduce multiple labels in the same tissue section): A table of substrate properties, a schematic of relative senstitivities of substrate formulations, and a table of pairwise substrate compatibility are also included.