Ready to use immunohistochemical secondary antibody kit (anti rabbit&mouse)

packaging: 5ml (can be used for 50 pieces)

applicable: primary antibody is from mouse or rabbit

chromogenic substrate: dab (benzidine)

reagent composition:

reagents not provided but required: primary antibody (from mouse or rabbit), 5% goat serum (abs933), pbst

this kit is the latest generation of non biotin detection system of immunohistochemical secondary antibody based on polymer technology, with stronger signal and simpler operation. Compared with the traditional immunohistochemical secondary antibody kit, it has three characteristics: first, due to the uniqueness of the polymer, the polymer molecule formed by the direct combination of enzyme molecules and secondary antibody IgG molecules is highly sensitive, effectively reducing the amount of primary antibody used, and the primary antibody reagent can be diluted 2-4 times; Second, because the use of biotin is avoided, the non-specific background color development caused by biotin can be avoided, and a clearer color development and cleaner background can be obtained; Third, rapid and shorter reagent incubation time

storage and validity period

2-8° C save. Each component can be stored for at least 12 months

operation steps:

note: all the steps are operated at room temperature. The reagent can be directly titrated in a drop bottle or with a pipette. One drop =30-40ul

1Dewaxed and hydrated tissue sections

2Wash 2-3 times in PBST for 5 minutes

3According to the special requirements of the applied primary antibody, the tissue sections were pretreated (antigen retrieval)

4Wash 2-3 times in PBST for 5 minutes

5Add 100ul endogenous peroxidase blocking agent was incubated for 10 min to block endogenous peroxidase to reduce nonspecific background staining

6Wash 2-3 times in PBST for 5 minutes

7Add 100ul of 5% to 10% normal goat serum and incubate for 20 minutes at room temperature. If the primary antibody is diluted in a buffer containing 5% to 10% normal goat serum, this step can be omitted

8Wash 2-3 times in PBST for 5 minutes

9Add primary antibody and incubate at room temperature or 37 ℃ for 30 min

10Wash 2-3 times in PBST for 5 minutes

11Add 100ul HRP enzyme labeled anti mouse & Rabbit secondary antibody polymer was incubated for 30 min   Note: this liquid is sensitive to light, and keep away from light

12Wash 2-3 times in PBST for 5 minutes

13Use freshly prepared dab chromogenic solution (Dilute with dab Dilution DAB substrate , ready to use) , incubate at room temperature for 5 minutes (the time can be adjusted according to the color development), and rinse the slide with tap water

14Add 100ulhematoxylin , incubate for 3 minutes (the time can be appropriately adjusted according to the color development), and rinse with tap water to return to blue

15Dehydrated, transparent, sealed