HRP Substrate-Chromogen (AEC) Kit

Description:
3-amino, 9 ethyl-carbazole (AEC) Chromogen

Presentation:
The use of 3-amino,9-ethyl-carbazole as a chromogen in conjunction with peroxidase-based immunohistochemical, immunocytochemical and in situ hybridization procedures, provides the red chromogen depositions at the reaction sites on tissue or cell specimens. The AEC kit contains buffer, chromogen and H2O2 substrate. All reagents are provided in convenient dropper bottles in concentrated form. AEC chromogen can be used in both manual and automated immunohistochemical systems.

The AEC depositions are alcohol soluble, therefore no organic solvents should be used after AEC staining. Red AEC signal can be counterstained with Biømeda’s Aqueous Hematoxylin (Cat. No. M10). AEC is incompatible with Permount or other organic solvent-based mounteng media. Therefore it requires aqueous mounting media like Biømeda's Crystal Mount (Cat. No. M02, M03).

Specificity:
These products are designed to produce a color deposition on tissue sections processed by immunocytochemical techniques using alkaline phosphatase or peroxidase as tracers. When using these products, it is very important to select the appropriate chromogen substrate for the enzyme tracer being used in the assay.
One mL of final working solution will yield approximately 5-6 tissue sections. The number of tests performed will vary depending on the application. Estimations are derived from use with the capillary-action based MicroProbe™ system, which draws 150-200 mL per test.
Each 20-tablet kit contains sufficient reagents to make 100 mL of final working solution. The 100 tablet chromogen-only kit will yield 500 mL of final working solution. All other kits contain sufficient reagents to make 300 mL of final working solution.
These products are designed to produce a color deposition on tissue sections processed by immunocytochemical techniques using alkaline phosphatase or peroxidase as tracers. When using these products, it is very important to select the appropriate chromogen substrate for the enzyme tracer being used in the assay.
One mL of final working solution will yield approximately 5-6 tissue sections. The number of tests performed will vary depending on the application. Estimations are derived from use with the capillary-action based MicroProbe™ system, which draws 150-200 mL per test.
Each 20-tablet kit contains sufficient reagents to make 100 mL of final working solution. The 100 tablet chromogen-only kit will yield 500 mL of final working solution. All other kits contain sufficient reagents to make 300 mL of final working solution.


Storage:
Refrigerate at 4°C. Do not freeze.

Notes:
Working Chromogen Preparation: Remove the AEC Chromogen kit from the
refrigerator 20 minutes before the preparation of the working chromogen solution.

1.    Transfer 5 mL of distilled water into supplied             plastic bottle.
2.    Add 2 drops of Concentrated Buffer Reagent.             Mix.
3.    Add one drop of Concentrated Chromogen
Reagent. Mix thoroughly.    
4. After chromogen dissolves, add one drop of
Concentrated Substrate Reagent. Mix well.
5.    Apply AEC working solution to the specimens.
Before applying this solution make sure to wash
the specimens extensively with buffer to remove
tracing reagent containing peroxidase.
6.    Incubate for 5-10 minutes at room temperature.         Wash specimens with distilled water and proceed         with the counterstaining steps.* For automated immunohistochemistry use corresponding number of drops of each reagent according to amount of water needed in the individual procedure.Use the working chromogen solution within 4-6 hours from the time of the addition of the Concentrated Chromogen. Discard unused reagent according to local and state regulations.AEC is a suspected carcinogen. Avoid contact with skin and clothes.