Product Introduction:

AEC substrate color reagent is an authoritative and classical method that can oxidize 3-amino-9-ethyl carbazole substrate to form red insoluble product through peroxidase catalysis, so as to identify peroxidase activity and determine the expression of target molecules. It is suitable for the immunological detection of peroxidase labeled protein of horseradish root (protein immunoimprinting, immunohistochemistry, and in situ hybridization, etc.). The product has stable performance, convenient operation, high sensitivity, clear color rendering and good repeatability.

3-amino-9-ethylcarbazole (3-amino-9-ethylcarbazole; AEC is the color substrate of Peroxidase. Its molecular formula is C14H14N2 and molecular weight is 210.3. Under the catalysis of peroxidase, the electron donor AEC substrate loses electrons under the oxidation of hydrogen peroxide and shows color change and deposition, forming a stable red insoluble product (soluble in organic reagents).

3-amino-9-ethylcarbazole (AEC) is the chromogenic substrate of Peroxidase. Under the catalysis of peroxidase, AEC was oxidized by hydrogen peroxide to form stable red precipitate. The red product is insoluble in water, but soluble in organic solvents. The sensitivity of the AEC substrate color development kit is greatly improved by adding an enhanced color development reagent. The product has the advantages of convenient operation, clear color rendering and good repeatability, and is suitable for enzyme-induced color development in IHC and Western Blot experiments of HRP system.

Instruction:

1. Preparation of working fluid:

Take 850ul sterile double steaming water, add 50ul solution A, 50ul solution B, 50ul solution C in turn, and mix evenly, that is, prepare AEC working liquid. If a larger volume of working liquid is needed, it can be scaled up accordingly. This solution must be used now, stored away from light after preparation, and used within half an hour. After expiration, please discard the remaining liquid.

2. Color development:

1) Color development of western blot membrane: The prepared working droplets are added to the western blot membrane (or the western blot membrane is immersed in the AEC working liquid), and the color development time is generally 10-30min. After the color development is complete, the membrane is immersed in distilled water to terminate the reaction.

2) Tissue section staining: Add appropriate amount of AEC working solution to the tissue section to ensure that the sample can be fully covered. Incubate at room temperature for 10-30 min, avoid light, until the color develops to the expected shade. The color development time can be controlled under the microscope. After color development, washing with distilled water can terminate the color development reaction.

3) For tissue sections or cell samples, after the color reaction is terminated, it can be re-dyed with other dyes. For the film, after the color reaction is terminated, it can be dried at room temperature and stored away from light.

Note:

1. Because the color product generated by AEC is fat soluble, it is not suitable for gradient alcohol dehydration after color development in IHC. Xylene transparent and neutral gum sealing tablets should be sealed with water-based sealing tablets.

2. According to the situation, optimize the experimental conditions and adjust the color development time to obtain the best color development effect.

3. AEC has certain toxicity, please take necessary protective measures during operation.

Note：Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.