Materials and Reagents

1. Sphero Protein A Polystyrene Particles (6-8 µm) (Spherotech, catalog number: PAP-60-5 )
   
2. Human anti-p24^Gag HIV-1 IIIB polyclonal antibodies (ImmunoDX, catalog number 2503 )
   
3. Normal Human IgG (Sigma-Aldrich, catalog number: I2511 )
   
4. p24^Gag recombinant protein (Abcam, catalog number: ab43037 )
   
5. Anti-p24^Gag KC57 clone conjugated to fluorescein isothiocyanate (FITC) or to phycoerythrin also called RD1 (Beckman Coulter, catalog numbers: 6604665 and 6604667 )
   
6. 1x phosphate-buffered saline (PBS) without calcium and magnesium (Corning, catalog number: 21-031-CV )
   
7. Bovine serum albumin (BSA) (Axenia Biologix, catalog number: S200 )
   
8. Triton X-100 (Thermo Fisher Scientific, catalog number: BP151-500 )
   
9. 16% paraformaldehyde (PFA)  solution (Electron Microscopy Sciences, catalog number: 15710 )
   
10. Fluorescence-activated cell sorting (FACS) staining buffer (see Recipes)
    

Equipment

1. Eppendorf centrifuge
   
2. Rotating mixer
   
3. 96-well V-bottom plate (optional) (Thermo Fisher Scientific, catalog number: 12-565-216 )
   
4. Thermo Fisher lids for 96-well microplates (Thermo Fisher Scientific, catalog number: 14-245-53A )
   
5. Plate sealers (VWR International, catalog number: 62402-921 )
   
6. Eppendorf tubes
   
7. Multichannel pipetman (optional)
   
8. Pipetman
   
9. Tips (1 ml, 200 µl, and 10 µl)
   
10. Reagent reservoirs (optional)
    
11. Tissue culture centrifuge for Eppendorf tubes or for 96 well plates (optional)
    
12. 37 °C incubator
    
13. Flow cytometer
    Note: The FLAQ requires a flow cytometer equipped with a blue laser excitation (488 nm) and one measurement parameter. The photomultiplicator tube (PMT) with a 525/50 nm or a 585/42 band pass filter is used for the detection of FITC or RD1 (phycoerythrin) signals, respectively.
    

Software

1. Forecyt (Intellicyt)
   
2. FlowJoX software (Tree Star) or other FACS analysis software
   

Procedure