Hematoxylin

Hematoxylin
Catalog No.
MYF07
Specifications
500 ml (MYF07-1)

Product Details

This reagent is suitable for immunohistochemical staining of formalin-fixed, paraffin-embedded human tissue sections, and is used for nuclear counterstaining during the assay.
Application
IHC-P
Form
Liquid
Shelf life
1 year
Storage conditions
Store at room temperature, protected from light
  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.