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IHC
on
DIS
CO
VER
Y U
LTR
A R
esea
rch
Inst
rum
ent
PD-L1 (SP142) from Spring Bioscience, Inc., with DISCOVERY HQ-HRP detection and ChromoMap DAB detection in non-small cell lung cancer (NSCLC)
Reagents Needed from Ventana
Name Catalog number Ordering code
DISCOVERY Anti-Rabbit HQ 760-4815 07017812001
DISCOVERY Anti-HQ HRP 760-4820 07017936001
DISCOVERY ChromoMap DAB Kit (RUO) 760-159 05266645001
DISCOVERY CC1 950-500 06414575001
Hematoxylin II 790-2208 05277965001
Bluing Reagent 760-2037 05266769001
Reagents Needed from Spring Bioscience, Inc.
Name Catalog number Dilution in protocol
PD-L1/CD274 (SP142) M4420 1: 30
IHC
with
DIS
CO
VER
Y H
Q-H
RP
dete
ctio
n
RUO DISCOVERY Universal Staining procedure for DISCOVERY ULTRA Research Instrument
VENTANA
Empowering | Innovation
www.roche.comwww.ventana.com
© 2014 Ventana Medical Systems, Inc.
VENTANA, VENTANA logo and DISCOVERY are trademarks of Roche. All other trademarks are the property of their respective owners. W5204-8 0814A
1. Deparaffinization [Selected]
2. Warmup Slide to [67 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 Reservoir [Selected]
8. Warmup Slide to [100 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. CC1 72 Min [Selected]
18. CC1 80 Min [Selected]
19. CC1 88 Min [Selected]
20. CC1 92 Min [Selected]
21. Inhibitor [Selected]
22. Inhibitor CM [Selected]
23. Apply One Drop of Inhibitor CM, and Incubate for [12 Minutes]
24. Antibody [Selected]
25. Disable heat for 1st Antibody [Selected]
26. Apply One Drop of [PREP KIT 1] (Antibody), and incubate for [1 hour]
27. Linking Antibody [Selected]
28. 2nd Antibody [Selected]
29. Warmup Slide to [36 Deg C] from Very Low Temperatures (2nd Antibody)
30. Apply One Drop of [Anti-Rabbit HQ] (Detection #1), and incubate for [0 HR 16 Min]
31. Enzyme conjugate [Selected]
32. Apply One Drop of [Anti-HQ HRP] (Conjugate #1), and Incubate for [12 Minutes]
33. DAB [Selected]
34. Counterstain [Selected]
35. Use RB for Counterstain [Selected]
36. Apply One Drop of [HEMATOXYLIN II] (Counterstain), and incubate for [8 minutes]
37. Post Counterstain [Selected]
38. Use RB for Post Counterstain [Selected]
39. Apply One Drop of [BLUING REAGENT] (Post Counterstain), and incubate for [4 minutes]