DAB Chromogen

Catalog No.
MYF16
Specifications
60 ml DAB substrate buffer + 3 ml chromogen (MYF16-1), 240 ml DAB substrate buffer + 12 ml chromogen (MYF16-2)

Product Details

This reagent is intended for immunohistochemical staining of formalin-fixed, paraffin-embedded human tissue sections, and is used for DAB chromogenic detection after secondary antibody binding.
    1. Soak tissue sections in two changes of fresh xylene for 10 min each, then in absolute ethanol, 95% ethanol, and 85% ethanol for 5 min each. Rinse gently in distilled water for 1 min.
    2. Place sections in Tris-EDTA (pH 9.0) antigen retrieval solution in a pressure cooker; heat until steaming and time for 2 min. After timing, allow to cool naturally for 5 min, then cool rapidly with water. Rinse gently in distilled water twice for 3 min each.
    3. Incubate sections in peroxidase blocking reagent (3% H2O2) for 10 min. Rinse gently in distilled water twice for 3 min each.
    4. Blot excess liquid around the tissue; circle the tissue edge 2–3 mm away with an IHC pen. Rinse gently in distilled water twice for 3 min each, then immerse in PBS buffer.
    5. Apply sufficient primary antibody working solution to cover the tissue completely. Incubate at room temperature in a humid chamber for 30–60 min. Wash with PBS 2–3 times and rinse gently twice for 3 min each.
    6. Apply sufficient secondary antibody working solution to cover the tissue completely. Incubate at room temperature in a humid chamber for 20 min. Wash with PBS 2–3 times and rinse gently twice for 3 min each.
    7. Apply sufficient DAB working solution to cover the tissue completely. Incubate at room temperature in a humid chamber for 5 min. Stop color development with distilled water.
    8. Counterstain in hematoxylin for 8–10 s. Rinse thoroughly with distilled water. Bluing in 1% lithium carbonate for 8–15 s, then rinse with distilled water.
    9. Dehydrate through 85% ethanol, 95% ethanol, and absolute ethanol for 3 min each, then in one change of fresh xylene for 5–10 min. Mount with neutral resin and examine under a microscope.
    Application
    IHC-P
    Form
    Liquid
    Shipping
    Biological refrigerated ice packs
    Storage conditions
    2–8°C
    Shelf life
    2 years
    Storage notes
    Store protected from light and heat