Table 1 mIF panels and staining conditions
Antibody | Clone | Source | Dilution | Fluorophore | |
|---|---|---|---|---|---|
1st panel | CD4 | 4B12 | ThermoFisher Scientific | 1:10 | Opal-650 |
CD8 | C8/144B | Agilent | 1:200 | Opal-480 | |
CD68 | KP1 | Agilent | 1:100 | Opal-540 | |
CD20 | L26 | Agilent | 1:400 | Opal-620 | |
FoxP3 | D2W8E | Cell Signalling (Danvers, MA, USA) | 1:50 | Opal-570 | |
Granzyme B | 11F1 | Leica Biosystems | 1:100 | Opal-520 | |
CD163 | 10D6 | Leica Biosystems | 1:150 | Opal-690 | |
Pan-cytokeratin (CK) | AE1/AE3 | Agilent | 1:100 | Opal-780 | |
Spectral DAPI | Akoya Bioscences | ||||
2nd panel | Neutrophil Elastase | NP57 | Agilent | 1:100 | Opal-570 |
CD44v3 | VFF-327-v3 | ThermoFisher Scientific | 1:40 | Opal-520 | |
CD44v6 | VFF-7 | ThermoFisher Scientific | 1:200 | Opal-480 | |
CD44v9 | RV3 | ThermoFisher Scientific | 1:50 | Opal-620 | |
CD56 | 123C3 | Agilent | 1:30 | Opal-540 | |
CD44s | F10-44-2 | Abcam (Cambridge, UK) | 1:200 | Opal-690 | |
Ki-67 | MM-1 | Leica Biosystems | RTU | Opal-650 | |
Pan-cytokeratin (CK) | AE1/AE3 | Agilent | 1:100 | Opal-780 | |
Spectral DAPI | Akoya Bioscences | ||||
3rd panel | ICAM-1 (CD54) | E3Q9N | Cell Signalling | 1:150 | Opal-540 |
RHAMM (CD168) | EPR4055 | Abcam | 1:100 | Opal-570 | |
Pan-cytokeratin (CK) | AE1/AE3 | Agilent | 1:100 | Opal-780 | |
Spectral DAPI | Akoya Bioscences |