Table 1.
Sequential IHC panel information.
| Cycle1 | Cycle2 | Cycle3 | Cycle4 | Cycle5 | Cycle6 | Cycle7 | Cycle8 | Cycle9 | Cycle10 | Cycle11 | Cycle12 | Cycle13 | Cycle14 | Cycle15 | |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| R1 | R2 | R5 | R7 | R9 | R11 | R13 | R15 | R17 | R19 | R21 | R23 | R25 | R27 | R29 | |
| Primary Ab | Hematoxylin | tBTK | PD1 | PD-L1 | ICOS | GATA-3 | CD66b | CD45 | T-bet | GzmB | CD68 | CD4 | Ki67 | SMA | p16 |
| Clone/Product# | D3H5 | NAT105 | E1L3N | SP98 | L50–823 | G10F5 | H130 | EPR9301 | GB7 | PG-M1 | SP35 | SP6 | ab5694 | E6H4 | |
| Vender | Cell Signaling | Abcam | Cell Signaling | LSBio | BD | eBioscience | Thermo Scientific | Abcam | BioRad | Abacam | Ventana | Sigma- Aldrich |
Abcam | Ventana | |
| Host sp | Rb IgG | Ms IgG | Rb IgG | Rb IgG | Ms IgG | Ms IgM | Ms IgG | Rb IgG | Ms IgG | Ms IgG | Rb IgG | Rb IgG | Rb IgG | Ms IgG | |
| Conc. | 1/50 | 1/50 | 1/100 | 1/25 | 1/100 | 1/600 | 1/100 | 1/100 | 1/200 | 1/50 | 1/8 | 1/1000 | 1/200 | Original | |
| Reaction | 1min | RT, 30min | RT, 30min | RT, 60 min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min |
| AEC | 40min | 40min | 40min | 40min | 40min | 20min | 40min | 20min | 20min | 20min | 20min | 20min | 20min | 10min | |
| Cycle2 | Cycle3 | Cycle4 | Cycle5 | Cycle6 | Cycle7 | Cycle8 | Cycle9 | Cycle10 | Cycle11 | Cycle12 | Cycle13 | Cycle14 | Cycle15 | ||
| Cycle1 | R3 | R6 | R8 | R10 | R12 | R14 | R16 | R18 | R20 | R22 | R24 | R26 | R28 | ||
| Primary Ab | DC-LAMP | EOMES | NKp46 | DC-SIGN | CSF1R | CD3 | CD8 | CD163 | MHC II | CD20 | Foxp3 | Tryptase | Pan-CK | ||
| Clone/Product# | 1010E1.01 | ab2283 | 195314 | DC-28 | SP211 | SP7 | SP16 | 10D6 | EPR11226 | SP32 | 236A/E7 | AA1 | AE1/AE3 | ||
| Vender | Novus Biological | EMD Millipore | R&D | Santa Cruz | Abcam | Thermo Scientific | Thermo Scientific | Thermo Scientific | Abcam | Sigma-Aldrich | eBioscience | Abcam | Abcam | ||
| Host sp | Rat IgG | Rb IgG | Ms IgG | Ms IgG | Rb IgG | Rb IgG | Rb IgG | Ms IgG | Rb IgG | Rb IgG | Ms IgG | Ms IgG | Ms IgG | ||
| Conc. | 1/100 | 1/1000 | 1/20 | 1/100 | 1/300 | 1/150 | 1/100 | 1/100 | 1/20000 | 1/2000 | 1/1000 | 1/200000 | 1/2000 | ||
| Reaction | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 30min | RT, 60min | RT, 30min | RT, 30min | RT, 30min | ||
| AEC | 30min | 20min | 20min | 20min | 20min | 20min | 20min | 20min | 10min | 20min | 20min | 20min | 10min |
R4 at Cycle2: Primary Ab, RORgt (Clone 6F3.1, EMD Millipore, Ms IgG, 1/200, RT, 30min), AEC 10min