Table 1.
Antibody panels
| Antigen | Clone | Isotype | Fluorochrome | Labeling strategy | Source of primary Ab |
|---|---|---|---|---|---|
| Ex vivo Ki-67/CD4 + T cells | |||||
| CD3 | PPT3 | IgG1 | PE | secondary antibodya | in house |
| CD4 | 74-12-4 | IgG2b | Alexa488 | secondary antibodyb | in house |
| CD8α | 11/295/33 | IgG2a | PE-Cy7 | secondary antibodyc | in house |
| CD27 | b30c7 | IgG1 | Alexa647 | directly conjugatedd | in house |
| CD45RC | 3a56 | IgG1 | PerCP-Cy5.5 | directly conjugatede | in house |
| SLA-DR | MSA3 | IgG2a | Qdot605 | directly conjugated | in housef |
| Ki-67 | B56 | IgG1 | V450 | directly conjugated | BD Biosciences |
| Ex vivo Ki-67/CD8β + T cells | |||||
| CD3 | PPT3 | IgG1 | PE | secondary antibodya | in house |
| CD8β | PG164A | IgG2a | Alexa488 | Zenon labeling kitg | VMRD |
| CD27 | b30c7 | IgG1 | Alexa647 | directly conjugatedd | in house |
| SLA-DR | MSA3 | IgG2a | Qdot605 | directly conjugated | in housef |
| Perforin | δ-G9 | IgG2b | PerCP-eFluor710 | directly conjugated | eBioscience |
| Ki-67 | B56 | IgG1 | V450 | directly conjugated | BD Biosciences |
| Triple cytokine staining/CD4 + T cells | |||||
| CD4 | 74-12-4 | IgG2b | Alexa488 | secondary antibodyb | in house |
| CD8α | 11/295/33 | IgG2a | PE-Cy7 | secondary antibodyc | in house |
| CD27 | b30c7 | IgG1 | BV421 | biotin-streptavidinh | in house |
| IFN-γ | P2G10 | IgG1 | PE | directly conjugated | BD Biosciences |
| TNF-α | MAb11 | IgG1 | BV605 | directly conjugated | BioLegend |
| IL-2 | A150D3F1 | IgG2a | APC | directly conjugatedi | Life Technologies |
| Triple cytokine staining/CD8β + T cells | |||||
| CD3 | BB23-8E6-8C8 | IgG2a | PE-Cy7 | directly conjugated | BD Biosciences |
| CD8β | PG164A | IgG2a | Alexa488 | Zenon labeling kitg | VMRD |
| CD27 | b30c7 | IgG1 | BV421 | biotin-streptavidinh | in house |
| IFN-γ | P2G10 | IgG1 | PE | directly conjugated | BD Biosciences |
| TNF-α | MAb11 | IgG1 | BV605 | directly conjugated | BioLegend |
| IL-2 | A150D3F1 | IgG2a | APC | directly conjugatedi | Life Technologies |
| In vitro expanded CD8β + T cells | |||||
| CD4 | 74-12-4 | IgG2b | PerCP-Cy5.5 | directly conjugated | BD Biosciences |
| CD8β | PG164A | IgG2a | Alexa488 | secondary antibodyj | VMRD |
| CD107a | 4E9/11 | IgG1 | Alexa647 | directly conjugated | AbD Serotec |
| Pan-γδ | PPT16 | IgG2b | PE-Cy7 | biotin-streptavidink | in house |
| IFN-γ | P2G10 | IgG1 | PE | directly conjugated | BD Biosciences |
| TNF-α | MAb11 | IgG1 | BV605 | directly conjugated | BioLegend |
aGoat anti-Mouse IgG1-PE, Southern Biotech.
bGoat anti-Mouse IgG2b-Alexa488, Life Technologies.
cGoat anti-Mouse IgG2a-PE-Cy7, Southern Biotech.
dAlexa Fluor-647 Protein Labeling Kit, Life Technologies.
eLightning-Link™ PerCP-Cy5.5 Tandem Conjugation Kit, Innova Biosciences.
fCustom conjugation by Life Technologies.
gIgG2a-Alexa488 Zenon labeling kit, Life Technologies.
hStreptavidin-BV421, BioLegend.
iLightning-Link™ APC Conjugation Kit, Innova Biosciences.
jGoat anti-Mouse IgG2a-Alexa488, Life Technologies.
kStreptavidin-PE-Cy7, eBioscience.