Whole Blood Staining of Human Monocyte Subsets for Flow Cytometry

Materials and Reagents

1. Antibodies
   1. Mouse anti-human CD56 FITC (BD Biosciences, catalog number: 340410 )
      
   2. Mouse anti-human CD2 FITC (BD Biosciences, catalog number: 555326 )
      
   3. Mouse anti-human CD19 FITC (BD Biosciences, catalog number: 555412 )
      
   4. Mouse anti-human CD14 PerCP (Life Technologies, Invitrogen™, catalog number: MHCD1431 )
      
   5. Mouse anti-human CD16 Pac Blue (BD Biosciences, catalog number: 558122 )
      
   6. Mouse anti-human HLA-DR APC-Cy7 (BioLegend, catalog number: 307617 )
      
   
   
2. Other materials
   1. Human blood
      
   2. 10x BD FACS Lysing Solution (BD Biosciences, catalog number: 349202 )
      
   3. 20% formaldehyde (Tousimis, catalog number: 1008A )
      
   4. Phosphate buffered saline (PBS) (Life Technologies, Gibco^®, catalog number: 20012-027 )
      

Equipment

1. Standard Bench-top Centrifuge
   
2. 5 ml polypropylene tubes (BD Biosciences, Falcon^®, catalog number: 352063 )
   
3. BD LSR II flow cytometer
   
4. Glass Whole Blood Tube with K3EDTA (BD Vacutainer^®, catalog number: 366450 )
   

Software

1. FlowJo (Tree Star)

Procedure