Product Introduction:

This kit is suitable for AP color development in immunohistochemical test of rabbit IgG with primary antibody. SABC is designed for immunohistochemical and other immune assays to show the distribution of antigens in tissues and cells. StreptAvidin (StreptAvidin) is a protein extracted from Streptomyces, like avidin, has a high affinity for biotin, avidin is an alkaline protein (IP=10), and can be modified to become a neutral protein. The isoelectric point of streptavidin is close to neutral, and the non-specific adsorption to tissues and cells is very low. The background of streptavidin based immunohistochemical method is very low. SABC stands for StreptAvidin - Biotin Complex. SABC can form a complex of about 100 peroxidase and about 50 streptavidin. A large number of enzymes will ensure that SABC is highly sensitive.

Experimental procedure: (Paraffin section as an example, for reference only)

1. Routine dewaxing of slices to water (three times xylene, three times ethanol).

2. Optional steps: a, heat repair antigen, b, enzyme digestion, c, skip this step and go directly to the next step.

3. Add 5%BSA sealer and leave at room temperature for 20 minutes. Discard excess liquid, do not wash.

4. Dilute the primary antibody with diluent in a certain proportion (the diluted primary antibody can be stored at 4℃ for one week), or you can purchase the antibody diluent separately. Add diluted primary resistance at 37 ℃ for about 1 hour or 20℃ for about 2 hours. Can also be 4℃ overnight. PBS (pH7.2-7.4) wash 2 minutes x 3 times. The dilution, incubation time and temperature of the primary antibody are directly related to the staining intensity and background. Specific dilution and incubation time refer to the manufacturer's manual of primary antibody.)

5. According to the dosage, the Bio-Sheep anti-Rabbit IgG concentrate was diluted by 1:100 into the working liquid (1ml diluent was mixed with 10ul Bio-sheep anti-rabbit IgG concentrate). This working liquid can be stored at 4℃ for one week). Drop Bio-Sheep Anti-Rabbit IgG working liquid,20-37℃ for 30 minutes. PBS (pH7.2-7.4) wash 2 minutes x 3 times.

6. According to the amount of use, the SBC-AP concentrate was diluted by 1:100 to the working liquid (1ml diluent was added to 10ul SBC-AP concentrate and mixed. This working liquid can be stored at 4℃ for one week). Drop ABC-AP working liquid at 20-37℃ for 30 minutes. PBS (pH7.2-7.4) wash 5 minutes x 4 times.

Seven. According to the dosage, add 40ul NBT solution to 1ml AP buffer, mix well, and then add 40ul BCIP solution. This working liquid is ready to use. Color development at room temperature, controlled reaction time under mirror. Distilled water washing terminates the reaction. * For frozen slices, after fixing, rinse twice with PBS and follow the second step above.

Note:

If the dyeing background is too high, after the SABC reaction and before AP color development, the slices are washed with PBST (pH7.2-7.4) added with 0.01-0.02% TWEEN-20 for 4 times, PBS for 2 times, and then AP color development. Because there are many immunohistochemical indicators and different specimens, this step is for reference only.

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