TintoFast CK AE1 & AE3 (AE1 & AE3), MMab

Presentation

1. 1. Embed the specimen in OCT inside a cryostat.
2. 2. Cut sections at 4-5 µm and mount on a positively charged glass slide such as the Bio SB Hydrophilic Plus Slides (BSB 7028) or in the lower third of the TintoDetector Cap Gap slides (BSB 7006).
3. 3. Air dry the slide at room temperature for 2 minutes and then incubate the slide at 60 °C for 3 minutes in an incubator or dry bath.
4. 4. Fix in 100% acetone or 10% NBF for 2 minutes at room temperature. NBF 10% produces better morphology.
5. 5. Rinse with distilled water and air dry the slides for another 2 minutes at room temperature.

1. 1. Transfer slides to ImmunoDNA washer, TBST or PBST buffer.
2. 2. For manual staining, perform antibody incubation at ambient temperature. For automated staining methods, perform antibody incubation according to instrument manufacturer’s instructions.
3. 3. Wash slides with ImmunoDNA washer, TBST, PBST or DI water.
4. 4. Continue IHC detection protocol. Wash slides between each step with ImmunoDNA washer, Tris or PBS Buffer solution.

1. 1. Do PIER. proleolytic digestion, with Mohs ImmunoDigestor for 1 min. Cool off
2. 2. Wash with Buffer (TBST or PBST)
3. 3. Incubate slides with Peroxidase Blocker for 30 seconds
4. 4. Wash with Buffer (TBST or PBST)
5. 5. Incubate CK for 4 min
6. 6. Wash with Buffer (TBST or PBST)
7. 7. Incubate with HRP Label for 3 min.
8. 8. Wash with Buffer (TBST or PBST)
9. 9. Prepare
   · DAB  Brown (1 drop of DAB chromogen in 1 ml of DAB Buffer; mix well)
   · or HRP Green (1 drop of HRP Green chromogen in 1 ml of HRP Green Buffer, mix well)
10. 10. Incubate with DAB or HRP Green for 2 min
11. 11. Wash with Buffer (TBST or PBST)
12. 12. Counterstain with Hematoxylin or Nuclear Fast Red for 30 seconds
13. 13. Wash with Buffer (TBST or PBST)
14. 14. Mount with AquaMounter or PermaMounter