Table 3.
Reagents, enzymes, chemicals, and solutionsa)
| Reagent | Manufacturer | Ordering Number |
|---|---|---|
| Chemicals & Solutions | ||
| RPMI 1640 | Thermo Scientific | 31870025 |
| Dulbecco’s Modified Eagle Medium (DMEM), high glucose | Thermo Scientific | 41965062 |
| Dulbeccós Phosphate Buffered Saline (PBS), no calcium, no magnesium | Thermo Scientific | 14190169 |
| l-Glutamine (200 mM) | Thermo Scientific | 25030-024 |
| Penicillin-Streptomycin (10,000 U/mL) | Thermo Scientific | 15140122 |
| 2-Mercaptoethanol (50 mM) | Thermo Scientific | 31350010 |
| Fetal bovine serum (FBS) | Gibco | 10270106 |
| UltraPure 0.5 M EDTA, pH8.0 | Thermo Scientific | 15575020 |
| Bovine Serum Albumin (BSA) | Sigma | A8806 |
| Trypan Blue solution | Sigma | T8154 |
| Recombinant Human Flt3-Ligand | Peprotech | 300-19 |
| Propidium Iodide (PI) - 1.0 mg/mL Solution in Water | Thermo Scientific | P3566 |
| Rainbow Calibration Particles | BD | 556286 |
| Antibodies | ||
| CD11c-APC (clone N418) | BD | 550261 |
| CD11c-PE (clone N418) | eBioscience | 12-0114-81 |
| Siglec-H-PE (clone 551) | BD | 129606 |
| MHC-II-BV650 (clone M5/114.15.2) | Biolegend | 107641 |
| CCR9-PEcy7 (clone eBioCW1.2) | eBioscience | 25-1991-82 |
| XCR1-APCcy7 (clone REA707) | Milteny | 130-111-375 |
| Sirp-α-PerCPeF710 (clone p84) | eBioscience | 46-1721-80 |
| Sirp-α-APC (clone p84) | Biolegend | 144013 |
| CD24a-FITC (clone M1/69) | eBioscience | 11-0242-81 |
| CD11b-eF450 (clone M1/70) | eBioscience | 48-0112-80 |
| B220-PerCP-cy5.5 (clone RA3-6B2) | eBioscience | 45-0452-80 |
a)
Reagents can be purchased from other vendors. For antibodies, alternative clones and fluorochromes can be used but should tested in advance.