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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:
- Deparaffinize the sections to water.
- Repair in repair fluid. Generally Tris-EDTA or citric acid repair solution.
- Add peroxidase blocker (usually 3% hydrogen peroxide) on the tissue and incubate for 15 minutes in the dark. Rinse with distilled water.
- Rinse with PBS three times, 5 minutes each time.
- 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.
- Shake off the blocking solution, add 50-100μL of the primary antibody working solution to each slice, overnight at 4°C.
- Rinse with PBS three times, 5 minutes each time.
- 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.
- Rinse with PBS three times, 5 minutes each time.
- 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.
- 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.
- 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.
- The slides are dehydrated with gradient alcohol (70-100%), transparent by xylene, and the slides are mounted with neutral gum.
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