Table 1.
Flow cytometry staining reagents
| Cell population | Primary antibody (mouse-anti-chicken) | Clone/isotype | Secondary antibody |
|---|---|---|---|
| NK cells | CD45-FITC1 | LT40/IgM | – |
| CD3-APC1 | CT3/IgG1 | – | |
| IL-2Rα-UNL2 | 28-4/IgG3 | Goat-anti-mouse-IgG3-PE1 | |
| 20E5-BIOT2 | IgG1 | Streptavidin (SA)-PercP6 | |
| T cells | CD3-PE1 | CT3/IgG1 | – |
| CD4-APC1 | CT4/IgG1 | – | |
| TCRγδ-FITC1 | TCR-1/IgG1 | – | |
| CD8α-UNL1 | EP72/IgG2b | Goat-anti-mouse-IgG2b-APC/Cy71 | |
| CD8β-BIOT1 | EP42/IgG2a | SA-PercP6 | |
| APCs | CD41/61-FITC4 | 11C3/IgG1 | – |
| Bu-1-AF6471 | AV20/IgG1 | – | |
| CD3-FITC1 | CT3/IgG1 | – | |
| CD4-PE/Cy71 | CT4/IgG1 | – | |
| MRC1LB-PE1 | KUL01/IgG1 | – | |
| CD11-biotin2 | 5C7/IgG1 | SA-Brilliant Violet (BV) 6057 | |
| MHC-II-UNL1 | Cia/IgM | Rat-anti-mouse-IgM-BV421 (RMM-1)7 | |
| *CHIR-AB1-UNL2 | 8D12/IgG2a | Rat-anti-mouse-IgG2a-PerCP/Cy5.5 (RMG2a-62)7 | |
| *CD40-UNL5 | AV79/IgG2a | Rat-anti-mouse-IgG2a-PerCP/Cy5.5 (RMG2a-62)7 | |
| *CD80-UNL5 | IAH:F864:DC7/IgG2a | Rat-anti-mouse-IgG2a-PerCP/Cy5.5 (RMG2a-62)7 | |
| Assay | |||
| CD107 | CD107a-APC3 | LEP-100 I 5G10/IgG1 | – |
| CD41/61-FITC4 | 11C3/IgG1 | – | |
| CD3-PE1 | CT3/IgG1 | – | |
| CD8α-UNL1 | EP72/IgG2b | Goat-anti-mouse-IgG2b-Alexa Fluor (AF) 7908 | |
| 28–4-UNL2 | IgG3 | Goat-anti-mouse-IgG3-APC/Cy71 | |
| 20E5-BIOT2 | IgG1 | SA-PercP6 | |
| IFNγ | CD3-PE1 | CT3/IgG1 | – |
| TCRγδ-FITC1 | TCR-1/IgG1 | – | |
| CD8α-UNL1 | EP72/IgG2b | Goat-anti-mouse-IgG2b-AF7908 | |
| 28-4-UNL2 | IgG3 | Goat-anti-mouse-IgG3-APC/Cy71 | |
| 20E5-BIOT2 | IgG1 | SA-PercP6 | |
| IFNγ-APC3 | MAb80/IgG1 | – | |
| T cell proliferation | CD3-FITC1 | CT3/IgG1 | – |
| CD4-PE/Cy71 | CT4/IgG1 | – | |
| CD8α-APC1 | CT8/IgG1 | – | |
| IL-2Rα-UNL2 | 28-4/IgG3 | Goat-anti-mouse-IgG3-PE1 | |
Three APC antibody panels were prepared each containing surface markers plus one out of three antibodies indicated (*).
Manufacturer: 1Southern Biotech, AL, USA.
2Purified antibody from hybridoma supernatant [44], kindly provided by Göbel, T.W., Ludwig Maximilian University, Germany.
3Developmental Studies Hybridoma Bank (DSHB), University of Iowa, IA, USA.
4Serotec, United Kingdom.
5Bio-Rad.
6BD Biosciences.
7Biolegend.
8Jackson ImmunoResearch Laboratories, PA, USA.