Specificity: anti-Mouse IgG (H+L), anti-Rabbit IgG (H+L)
Chromogen/Substrate: None provided
TP-060-HL: 60mL Ultra V Block (TA-060-UB), 60mL Biotinylated Goat Anti-Polyvalent (TP-060-BN), 60mL Streptavidin Peroxidase (TS-060-HR)
TP-125-HL: 125mL Ultra V Block (TA-125-UB), 125mL Biotinylated Goat Anti-Polyvalent (TP-125-BN), 125mL Streptavidin Peroxidase (TS-125-HR)
UltraVision detection system detects a specific antibody bound to an antigen in tissue sections. The specific antibody is located by a biotin-conjugated secondary antibody. This step is followed by the addition of a streptavidin-enzyme conjugate that binds to the biotin present on the secondary antibody. The specific antibody, secondary antibody, and streptavidin-enzyme complex is then visualized with an appropriate substrate/chromogen.
1. Deparaffinize and rehydrate tissue section.
2. To reduce nonspecific background staining due to endogenous peroxidase, incubate slide in hydrogen peroxide for 10-15 minutes.
3. Wash 2 times in buffer.
4. If required, incubate tissue in digestive enzyme (or appropriate pretreatment).
5. Wash 4 times in buffer.
6. (Optional) Apply Ultra V Block and incubate for 5 minutes at room temperature to block nonspecific background staining.
NOTE: Do not exceed 10 minutes or there may be a reduction in desired stain.
7. Rinse (Optional).
8. Apply primary antibody and incubate according to manufacturer's protocol.
9. Wash 4 times in buffer.
10. Apply Biotinylated Goat Anti-Polyvalent and incubate for 10 minutes at room temperature.
11. Wash 4 times in buffer.
12. Apply Streptavidin Peroxidase and incubate for 10 minutes at room temperature.
13. Rinse 4 times in buffer.
14. Incubate with peroxidase-compatible chromogen of choice according to manufacturer's recommendations.
15. Counterstain and cover slip.
The specificity and sensitivity of antigen detection is dependent on the specific primary antibody used.