Monosan Permanent AEC Kit, 200 ml

Intended use: Permanent AEC Kit is intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). AEC (3-Amino-9-ethylcarbazol) leads to the formation of a red-brown precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy. Permanent AEC Kit is intended for research use only.

Reagents provided: 5.5 ml Reagent 1 3 ml Reagent 2 3 ml Reagent 3 (Chromogen)4.5 ml Reagent 4 (H2O2)1 Dilution Vial

Storage and handling: The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution should be prepared freshly at the day of use. Excess working solution should be disposed. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.

Reagent preparation: 1) Pipette 5 ml distilled or deionised water into the provided dilution vial. 2) Add 3 drops buffer concentrate (Reagent 1). Mix thoroughly. 3) Add 2 drops Reagent 2. Mix thoroughly. 4) Add 2 drops AEC chromogen (Reagent 3). Mix thoroughly. 5) Add 2 drops H2O2 substrate (Reagent 4). Mix thoroughly. This working solution is stable for at least 16 hours if stored at 2-8°C in a dark place.

Procedure: 1) Apply the Permanent AEC working solution onto the slide. Incubate for 5-15 minutes. (Incubation time can be extended, if desired.) 2) Rinse with distilled or deionised H2O. 3) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 4) Rinse with distilled or deionised H2O. 5) Blueing in tap water for at least 5 minutes. 6) Dehydrate through a graded series of ethanol and clear in xylene. Mount with a permanent mounting medium.

Expected results: During the reaction of the substrate with horse radish peroxidase in presence of the chromogen AEC, a red-brown precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in aqueous and organic solvents and can be observed by light microscopy.

References: Elias JM Immunohistopathology – A practical Approach to Diagnosis ASCP Press 2003/Nadji M and Morales AR Ann N.Y. Acad Sci 420:134-139, 1983

• SKU: SANMON-APP173
• Manufacturer: Sanbio / Monosan
• Manufacturer SKU: MON-APP173
• Green Labware: No
• Package Unit: 2000 Tests
• Quantity Unit: PAK
• Application: Immunofluorescence, Immunohistochemistry (frozen), Immunohistochemistry (paraffin)