ihc on discovery ultra research instrument · discovery anti-hq hrp 760-4820 7017936001 discovery...
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PTEN (D301) (SP227) from Spring Bioscience, Inc., with DISCOVERY HQ-HRP Detection and DISCOVERY Purple Chromogen in prostate carcinoma
Reagents Needed from Ventana
Name Catalog number Ordering code
DISCOVERY Anti-Rabbit HQ 760-4815 7017812001
DISCOVERY Anti-HQ HRP 760-4820 7017936001
DISCOVERY Inhibitor 760-4840 7017944001
DISCOVERY Purple kit (RUO) 760-229 7053983001
DISCOVERY CC1 950-500 6414575001
Hematoxylin II 790-2208 5277965001
Bluing Reagent 760-2037 5266769001
Reagents Needed from Spring Bioscience
Name Part number Dilution in protocol
Cleaved PTEN (D301) (SP227) M5274 1:300
Antibody Diluent ADS-125IHC
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en1. Deparaffinization [Selected]
2. Warmup Slide to [69 Deg C], and incubate for [8 Minutes] (Cycle 1)
3. Incubate for [8 Minutes] (Cycle2)
4. Incubate for [8 Minutes] (Cycle3)
5. Pretreatment [Selected]
6. Cell Conditioning [Selected]
7. CC1 Resivoir [Selected]
8. Warmup Slide to [93 Deg C], and incubate for 4 Minutes (Cell Conditioner #1)
9. CC1 8 Min [Selected]
10. CC1 16 Min [Selected]
11. CC1 24 Min [Selected]
12. CC1 32 Min [Selected]
13. CC1 40 Min [Selected]
14. CC1 48 Min [Selected]
15. CC1 56 Min [Selected]
16. CC1 64 Min [Selected]
17. Dual Sequence [Selected]
18. DS Inhibitor [Selected]
19. Neutralize [Selected]
20. DS Antibody [Selected]
21. DS Antibody Manual Application [Selected]
22. Warmup Slide to [36 Deg C] from Very Low Temperatures (DS Primary Antibody)
23. Hand Apply (DS Primary Antibody), and incubate for [40 minutes]
24. DS Linking Antibody [Selected]
25. DS 2nd Antibody [Selected]
26. Warmup Slide to [36 Deg C] from Very Low Temperatures (DS Primary Antibody)
27. Apply One Drop of [Anti-Rabbit HQ] (Detection #5), and incubate for [0 Hr 12 Min]
28. DS Enzyme conjugate [Selected]
29. Apply One Drop of [Anti-HQ HRP] (Conjugate #2), and incubate for [12 Minutes]
30. DS DISCOVERY Purple [Selected]
31. Apply One Drop of DISC H2O2 P, and incubate for [1 Hour]
32. Counterstain [Selected]
33. Use RB for Counterstain [Selected]
34. Apply One Drop of [HEMATOXYLIN II] (Counterstain), and incubate for [4 minutes]
35. Post Counterstain [Selected]
36. Use RB for Post Counterstain [Selected]
37. Apply One Drop of [BLUING REAGENT] (Post Counterstain), and incubate for [4 minutes]
RUO DISCOVERY Universal Staining procedure for DISCOVERY ULTRA Research Instrument
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