Specificity: anti-Mouse IgG (H+L), anti-Rabbit IgG (H+L)
Enzyme: Alkaline Phosphatase
Chromogen/Substrate: Fast-Red/Naphthol Phosphate Buffer
TP-015-AF: 15mL Ultra V Block (TA-015-UB), 15mL Biotinylated Goat Anti-Polyvalent (TP-015-BN), 15mL Streptavidin Alkaline Phosphatase (TS-015-AP), 15mL Naphthol Phosphate Substrate (TA-015-AFS), three Fast-Red Tablets (TA-003-AFC)
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. Wash 2 times in buffer.
3. If required, incubate tissue in digestive enzyme (or appropriate pretreatment).
4. Wash 4 times in buffer.
5. (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.
6. Rinse (Optional).
7. Apply primary antibody and incubate according to manufacturer's protocol.
8. Wash 4 times in buffer.
9. Apply Biotinylated Goat Anti-Polyvalent and incubate for 10 minutes at room temperature. (Some users have experienced better staining when incubating for 20 minutes)
10. Wash 4 times in buffer.
11. Apply Streptavidin Alkaline Phosphatase and incubate for 10 minutes at room temperature. (Some users have experienced better staining when incubating for 20 minutes)
12. Rinse 4 times in buffer.
13. Add 1 Fast Red tablet to 5mL of Naphthol Phosphate Substrate and shake until tablet is dissolved. Apply to tissue section and incubate for 10-20 minutes or until desired stain intensity is achieved.
WARNING: Handle with care and dispose of according to all regulations.
14. Counterstain and cover slip.
The specificity and sensitivity of antigen detection is dependent on the specific primary antibody used.