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AEC, 50X (3-amino-9-ethylcarbazole); for HRP-based detection systems; provides red chromogen depositions. Precipitable peroxidase substrate for immunohistochemical (IHC) staining techniques. In the presence of horseradish peroxidase (HRP), produces a water-insoluble red oxidation product. Although not as sensitive as diaminobenzidine (DAB, 50X;
sc-24982), the red deposit formed at the sites of AEC oxidation make this substrate useful for double labeling. Aqueous-based mounting medium and counterstains must be used for IHC applications since oxidized AEC is soluble in alcohols and xylene (e.g., Crystal Mounting Medium; sc-24943). Use with AEC Substrate Buffer (
sc-36873).
• Prepare samples and incubate with antibodies and HRP-labeled probes, follow with all necessary wash steps.
• Prepare 1X AEC Substrate Buffer (dilute AEC Substrate Buffer, 10X;
sc-36873).
• Dilute one part AEC, 50X with 50 parts 1X AEC Substrate Buffer to make 1X AEC Substrate Solution. Mix well.
• Cover tissue sections with 1X AEC Substrate Solution and incubate 10-30 min. at RT.
• Rinse thoroughly with distilled water. If desired, follow with an appropriate aqueous-based counterstain.
• Mount tissue samples with an aqueous-based mounting medium (e.g., Crystal Mounting Medium; sc-24943)
• Unused AEC solution can be discarded according to city, county, state, province or country’s regulations.