Table 5.
Multiplexed immunofluorescence antibodies and their targets.
| Target | Antibody clone | Source | Dilution and antigen retrieval | Opal fluorophores |
|---|---|---|---|---|
| CD3 | F7.2.38 | DAKO M7254 | 1:100, E-AR | Opal 780 |
| CD8 | C8/144B |
Invitrogen MA5-13473 |
1:100, E-AR | Opal 690 |
| CD20 | EP459Y | Abcam ab78237 | 1:200, C-AR | Opal 620 |
| CD68 | CD68/684 | Abcam ab201340 | 1:4000, C-AR | Opal 480 |
| PD-1 | EPR4877(2) | Abcam ab137132 | 1:50, C-AR | Opal 520 |
| PD-L1 | E1L3N | Cell signalling #13684 | 1:25, E-AR | Opal 570 |
| FOXP3 | 236A/E7 | Abcam ab20034 | 1:1000, E-AR | Opal 540 |
| Pan-CK | AE1/AE3 | Abcam ab27988 | 1:3000, C-AR | Opal 650 |
C-AR, citrate antigen retrieval at pH 6.0; E-AR, Ethylenediaminetetraacetic acid (EDTA) antigen retrieval at pH 9.0; Pan-CK, pan-cytokeratin; PD-1, programmed cell death protein-1; PD-L1, programmed death-ligand 1.