AEC Chromogen/Substrate

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Product Details

Summary
Applications IHC, IHC-Fr, IHC-P

Order Details

AEC Chromogen/Substrate Summary

Specificity
Peroxidase reacts with 3% Hydrogen Peroxide Substrate to degrade it, which in turn reacts with AEC to precipitate it at the positive sites giving red brown colored end product.

Applications/Dilutions

Dilutions
  • Immunohistochemistry 1:10-1:500
Application Notes
To use as substrate/chromogen in conjunction with peroxidase based immunostaining systems.
Note: The working chromogen solution is stable for 6 hours. Any solution not used after this period should be discarded.
1. Take 5 ml of distilled or de-ionized water in a test tube.
2. Add two drops of concentrated buffer and mix.
3. Add two drops of concentrated AEC Chromogen and mix.
4. Add two drops of 3% H2O2 substrate solution and mix.

Procedure:
1. Once tissue sections have been incubated with peroxidase, wash them with buffer thoroughly.
2. Wipe the class to remove excess buffer and add enough drops of the working AEC solution to cover the tissue sections.
3. Incubate for 10-20 mintues at room temperature. For the best results, look under the microscope for the signal development. Once desired signal to noise ratio is achieved, stop the reaction by washing slides in wash buffer.

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.

Notes

Kit Components: 3mL concentrated AEC Buffer, 3mL concentrated AEC Chromogen and 3 mL of 3% Hydrogen Peroxide Substrate.

Background

Even though DAB is much more sensitive chromogen compared to amino ethyl carbazol (AEC) because of its carcinogenic nature, some labs avoid using DAB. To resolve this problem, we have designed an AEC chromogen in liquid format. AEC is a chromogen of choice when performing immunoperoxidase staining. It has been well accepted amongst the pathologist because of its relatively less toxic nature compared to DAP. It produces a red colored end product at the positive sites which gives a good contrast with blue hematoxylin counter stain. Specimens stained in AEC cannot be dehydrated in ethanol and hence should be mounted in aqueous based mounting medium.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Support products are guaranteed for 6 months from date of receipt.

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