Specificity: anti-Mouse IgG (H+L), anti-Rabbit IgG (H+L)
Chromogen/Substrate: Diaminobenzidine (DAB)
TP-015-HD: 15mL Hydrogen Peroxide Block (TA-015-HP), 15mL Ultra V Block (TA-015-UB), 15mL Biotinylated Goat Anti-Polyvalent (TP-015-BN), 15mL Streptavidin Peroxidase (TS-015-HR), 15mL DAB Plus Substrate (TA-015-HSX), 1mL DAB PlusChromogen (TA-001-HCX)
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 Block 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. Add 1 drop (40μL) DAB Plus Chromogen to 2mL of DAB Plus Substrate, mix by swirling and apply to tissue. Incubate for 5-15 minutes, depending on the desired stain intensity.
WARNING: DAB is a suspected carcinogen. Handle with care and dispose of according to all regulations.
15. Counterstain and cover slip using a permanent mounting media.
The specificity and sensitivity of antigen detection is dependent on the specific primary antibody used.