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G1213-100UL Goat Anti-rabbit Secondary Antibody for IHC Western Blot

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G1213-100UL Goat Anti-rabbit Secondary Antibody for IHC Western Blot

This product is a particularly sensitive two-step immunohistochemistry kit when used in conjunction with the DAB chromogenic kit, which is suitable for rabbit-derived primary antibodies. This secondary antibody is a molecule formed by combining horseradish peroxidase and goat anti-rabbit IgG. The macromolecule after this molecule is combined with the rabbit-derived primary antibody (reacted with the antigen first) is labeled with horseradish peroxidase, and then React with DAB to obtain a brown-yellow precipitate. Since the system does not contain biotin and streptavidin, it is not interfered by endogenous biotin, the stained background is very clear.

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Technical Specification

Storage conditions:

Wet ice transportation ; store at -20°C, valid for 12 months.

 

Instructions:

  1. Deparaffinize the sections to water.
  2. Repair in repair fluid. Generally Tris-EDTA or citric acid repair solution.
  3. Add peroxidase blocker (usually 3% hydrogen peroxide) on the tissue and incubate for 15 minutes in the dark. Rinse with distilled water.
  4. Rinse with PBS three times, 5 minutes each time.
  5. Incubate 3% BSA (pH 7.2-7.4 PBS buffer solution) or normal goat serum working solution on the tissue for 30 minutes to block non-specific binding sites.
  6. Shake off the blocking solution, add 50-100μL of the primary antibody working solution to each slice, overnight at 4°C.
  7. Rinse with PBS three times, 5 minutes each time.
  8. Add 50-100 μL of the secondary antibody working solution to each slide (diluted in the ratio of 1:200 between the two antigen solution G1213 and pH7.2-7.4 PBST), and incubate at room temperature for 30-60 minutes.
  9. Rinse with PBS three times, 5 minutes each time.
  10. Preparation of DAB working solution: add 20μL of 50×DAB stock solution (G1215-2) to every 1 mL of DAB diluent (G1215-1) and mix well for later use. It can also be used after proper dilution according to the test results.
  11. Add 50-100 μL of freshly prepared DAB working solution to each slice, and incubate at room temperature. The microscope controls the color development time.
  12. After the color development is completed, rinse with distilled water or tap water. Hematoxylin counterstaining, differentiation with 0.1% hydrochloric acid, washing with tap water to return to blue.
  13. The slides are dehydrated with gradient alcohol (70-100%), transparent by xylene, and the slides are mounted with neutral gum.
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