AEC Peroxidase Substrate Solution

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Tony Rook
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AEC Peroxidase Substrate Solution

Please find the following media preparation method:

AEC Peroxidase Substrate Solution -
AEC (3-amino-9-ethylcarbazole) Peroxidase Substrate Solution Red

Final Dilution:

0.05% AEC - 0.015% H2O2 in 0.05M Acetate Buffer,
pH 5.5

Stock Solutions:

1% AEC (20x) in 100% DMF (N, N Dimethylformamide):

Dissolve 0.1 g of AEC in 10 ml of 100% DMF and store this
solution at 4 °C.

0.3% H2O2 (20x) in Distilled Water:

Add 100 ul of 30% H2O2 in 10 ml distilled water and store
at 4 °C.

1M Acetate Buffer (20x), pH 5.5:

Sodium acetate, trihydrate (MW 136.1) ------ 13.6 g

Distilled water ------------------------------ 100 ml

Mix to dissolve and adjust pH to 5.5 using concentrated
HCl or glacial acetic acid.

0.05M Acetate Buffer (1x), pH 5.5:

1M Acetate buffer --------------------------- 1 ml

Distilled water ------------------------------- 19 ml

Mix well and adjust pH if necessoary.

Working Solution:

Add 5 drops of 1% AEC to 5 ml of 0.05M Acetate Buffer,
pH5.5 and mix well.

Add 5 drops of 0.3% H2O2 and mix well.

Incubate sections for 10-15 minutes at room temperature.

Note: AEC color products can be washed off with ethanol/alcohol solution. So slides must be coverslipped using aqueous mounting media.

Link Reference:
http://www.ihcworld.com/_protocols/chromogen_substrates/POD_AEC_red.htm