Table 1. Summary of antibody reagents used for identification of IgG+ antigen-specific mouse B cells.
| Marker/Reagent | Cell type | Fluorophore |
|---|---|---|
| B220 (CD45R) | B-cells | MACS bead |
| CD19 | B-cells | AF700 |
| IgG | Memory B-cells | BV421 |
| Antigen | Ag specific B-cell | PE |
| Antigen | Ag specific B-cell | APC |
| IgM | Naïve B-cells | PE-Cy7 |
| IgD | Naïve B-cells | APC-Cy7 |
| CD4 | T-cells | FITC |
| CD8 | T-cells | FITC |
| GR-1 | Neutrophils | FITC |
| F4/80 | Macrophages | FITC |
| 7AAD | Dead cells |
An optimised panel of fluorophore-labelled reagents was developed to maximise the potential of identifying the IgG+ antigen-specific subset of B cells. Both B cell-specific and non-B cell specific (dump channel) reagents were employed.