Immunophenotyping of dendritic cells from human whole blood using flow cytometry

Miltenyi Biotec-tested panel 10 (MBTP 10)

A whole blood sample obtained from a healthy donor was analyzed for dendritic cell subsets.


This protocol includes detailed instructions for preparing necessary reagents and materials and then carrying out each step.


The following is a listing of reagents, instruments, and consumables needed for each step of this protocol. These products are for research use only.

Antibody panel details

SpecificityClonePurposeFluorochromeDetection filter (nm) (laser)
CD45REA747LeukocytesVioGreen™525/50 (violet)
CD303 (BDCA-2)REA693Plasmacytoid dendritic cellsPE585/40 (blue)
CD14REA599Cell type exclusionFITC525/50 (blue)
CD20REA780Cell type exclusionFITC525/50 (blue)
CD123REA918Plasmacytoid dendritic cellsAPC-Vio® 770750LP (red)
CD141 (BDCA-3)REA674Conventional dendritic cells 1APC655-730 (red)
CD1c (BDCA-1)REA694Conventional dendritic cells 2PE-Vio 770750LP (blue)
FCεR1αCRA1Conventional dendritic cells 2VioBlue®450/50 (violet)
7-AAD Staining Solution-Viability-655-730 (blue)

Additional reagents used

  • Phosphate-buffered saline (PBS), pH 7.2
  • PEB buffer: PBS, pH 7.2, 0.5% bovine serum albumin (BSA), and 2 mM EDTA.
  • Red Blood Cell Lysis Solution (# 130-094-183)
  • Flow cytometer, for example, MACSQuant® Analyzer 10 (# 130-096-343) or MACSQuant X (# 130-105-100)
  • (Optional) MACSQuant Calibration Beads (# 130-093-607)
  • (Optional) MACS® Comp Bead Kit, anti-REA (# 130-104-693)
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