The ImmunoAtlas and Abcam teams have collaborated and designed a panel of seven antibodies (α-CD8α, α-Foxp3, α-Granzyme B, α-PD-L1, α-PD-1, α-Cytokeratin, α-EpCAM; Abcam), which mark several phenotypes essential to immunotherapy responses. The panel enables the identification and quantification of different immune subpopulations such as cytotoxic T-cells, regulatory T cells, etc within the tumor microenvironment. Breast cancer tissue array samples from 70 patients (normal, stage I to III) were stained with the markers (refer table) and multiplex IHC kit (Opal). All the images were acquired using a multispectral imaging system (Vectra® Polaris™).
Markers |
Host species |
Clone |
Ab Dilution |
Supplier |
Product Cat # |
Fluorophore |
DAPI |
|
|
|
Akoya Biosciences |
FP1490 |
|
Anti-PD-L1 |
Rabbit |
Monoclonal (CAL10) |
1:750 |
Abcam |
ab251611 |
Opal 520 |
Anti-Granzyme B |
Rabbit |
Monoclonal (EPR20129-217) |
1:250 |
Abcam |
ab219803 |
Opal 540 |
Anti-PD1 |
Rabbit |
Monoclonal (CAL20) |
1:750 |
Abcam |
ab251613 |
Opal 570 |
Anti-pan Cytokeratin |
Mouse |
Monoclonal (C-11) |
0.5ug/ml |
Abcam |
ab264485 |
Opal 620 |
Anti-EpCAM |
Rabbit |
Monoclonal (EPR20532-225) |
1:1250 |
Abcam |
ab225894 |
Opal 620 |
Anti-CD8α |
Rabbit |
Monoclonal (CAL66) |
1:1500 |
Abcam |
ab251596 |
Opal 650 |
Anti-FOXP3 |
Mouse |
Monoclonal (236A/E7) |
10ug/ml |
Abcam |
ab96048 |
Opal 690 |
|