MaxVisionTM HRP Immunohistochemistry Staining Procedure
Ready-to-use Rapid Immunohistochemistry MaxVisionTMThe kit is based on polymer technology, where horseradish peroxidase is conjugated with anti-mouse/and anti-rabbit IgG molecules or anti-goat IgG molecules onto a polymer to form polymer molecules. The staining procedure is as follows:
1. Deparaffinize and hydrate paraffin sections, rinse with tap water;
2. Perform appropriate antigen retrieval on the tissue according to the requirements of the primary antibody;
3. If necessary (for tissues with high endogenous peroxidase content), apply peroxidase blocking reagent to the sections, incubate at room temperature for 10 minutes, rinse with PBS three times, 3 minutes each time (3 × 3 minutes);
4. Remove PBS, apply the primary antibody to the sections, incubate at room temperature for 60 minutes or at 4°C overnight, rinse with PBS 3 × 3 minutes;
5. Remove PBS, apply MaxVisionTM/HRP reagent to the sections, incubate at room temperature for 15 minutes, rinse with PBS 3 × 3 minutes;
6. Remove PBS, apply freshly prepared enhanced DAB chromogenic reagent to the sections for color development;
7. Rinse with tap water to stop color development, counterstain with hematoxylin, (if necessary, differentiate with 1% hydrochloric acid ethanol), return to blue with PBS. If DAB is used for color development, dehydrate the sections through a graded ethanol series, clear with xylene, and mount with neutral balsam; if AEC is used for color development, the sections cannot be dehydrated with ethanol and should be directly mounted with an aqueous mounting medium.
1. Deparaffinize and hydrate paraffin sections, rinse with tap water;
2. Perform appropriate antigen retrieval on the tissue according to the requirements of the primary antibody;
3. If necessary (for tissues with high endogenous peroxidase content), apply peroxidase blocking reagent to the sections, incubate at room temperature for 10 minutes, rinse with PBS three times, 3 minutes each time (3 × 3 minutes);
4. Remove PBS, apply the primary antibody to the sections, incubate at room temperature for 60 minutes or at 4°C overnight, rinse with PBS 3 × 3 minutes;
5. Remove PBS, apply MaxVisionTM/HRP reagent to the sections, incubate at room temperature for 15 minutes, rinse with PBS 3 × 3 minutes;
6. Remove PBS, apply freshly prepared enhanced DAB chromogenic reagent to the sections for color development;
7. Rinse with tap water to stop color development, counterstain with hematoxylin, (if necessary, differentiate with 1% hydrochloric acid ethanol), return to blue with PBS. If DAB is used for color development, dehydrate the sections through a graded ethanol series, clear with xylene, and mount with neutral balsam; if AEC is used for color development, the sections cannot be dehydrated with ethanol and should be directly mounted with an aqueous mounting medium.