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Table 1 Overview of the marker panels of primary monoclonal antibodies and 7-AAD to immunophenotype equine MSC
| Subset | Marker | Company | Clone | Sec Ab | Dilution | Positive control | |
|---|---|---|---|---|---|---|---|
| Multicolor FCM | 1 | CD29-Alexa488+ | Biolegend | TS2/16 | 1:20 | MNCs | |
| MHC-II− | Serotec | CVS20 | RPE | 1:50 | MNCs | ||
| 7-AAD− | Calbiochem | ||||||
| CD44-APC+ | BD | IM7 | 1:20 | MNCs | |||
| 2 | CD105-RPE+ | Serotec | SN6 | 1:10 | Endothelial cells | ||
| 7-AAD− | Calbiochem | ||||||
| CD90+ | VMRD | DH24A | Alexa647 | 1:100 | MNCs | ||
| 3 | CD45-Alexa488− | Serotec | F10-89-4 | 1:5 | MNCs | ||
| CD73+ | Abcam | 10f1 | RPE | 1:5 | Lymphocytes | ||
| 7-AAD− | Calbiochem | ||||||
| 4 | Monocyte-Alexa488− | Serotec | MAC387 | 1:2.5 | MNCs | ||
| CD79α-Alexa647− | Serotec | HM57 | 1:2.5 | MNCs | |||
| Single-color FCM | MHC-I+ | VMRD | PT85A | RPE | 1:66 | MNCs | |
| Secondary Ab | 1 and 3 | Sheep anti-mouse IgG-RPE | Sigma-Aldrich | 1:20 | |||
| 2 | Goat anti-mouse IgG-Alexa647 | Invitrogen Corporation | 1:200 | ||||
| Isotype controls | 1 and 4 | Mouse IgG1-Alexa488 | Biolegend | 1:20 | |||
| 1-3 | Mouse IgG1-RPE | Biolegend | 1:10 | ||||
| 1 | Rat IgG2b-APC | Biolegend | 1:20 | ||||
| 2 | Mouse IgM | BD | Alexa647 | 1:50 | |||
| 3 | Mouse IgG2-Alexa488 | Biolegend | 1:20 | ||||
| 4 | Mouse IgG1-Alexa647 | Biolegend | 1:100 |
