Anti p24 kc57 fitc antibody

The following antibodies were used at the recommended dilution of 0.25 µg antibody/million cells: CD3 (SK7), CD4 (SK3), CD16 (3GB), CD56 (B159), CD69 (FN50). Cell surface staining for CD69 activation was carried on CD56⁺/CD3⁻ gated NK cells with gates set upon unstimulated control cells. For intracellular staining of the HIV-1 p24 gag protein, CD4⁺ T cells were permeabilized with the Cytofix/Cytoperm kit (BD Pharmingen) as described by the manufacturer and stained with the anti-p24 KC57 FITC antibody (Beckman Coulter, CA). Samples were collected on a LSRII Cytometer (BD) and were analyzed with FlowJo software (Tree Star Incorporated, Ashland OR).

All antibodies were obtained from BD Biosciences (San Jose, California, USA) unless otherwise noted and used at the recommended dilution of 0.25 μg antibody/million cells. Cell surface staining for the CD69 activation marker and the CD16 FcγIII receptor was carried on CD56⁺/ CD3⁻ gated NK cells with gates set upon isotype control. Cell surface staining for gp120 on uninfected, HIV-1 infected, or gp120-coated CD4⁺ T cells was carried out using a 1 : 1000 dilution or plasma from a representative HIV-1-infected elite controller patient (viral load <50 copies/ml, ART-naïve) followed by detection with a phycoerythrin (PE)-conjugated goat secondary antibody to the Fc portion of human IgG. For intracellular staining of the HIV-1 p24 gag protein, CD4⁺ primary T cells were permeabilized with the Cytofix/Cytoperm kit (BD Biosciences) and stained with the antip24 KC57 FITC antibody (Beckman Coulter, Brea, California, USA) as described by the manufacturer. Samples were collected on a LSRII Cytometer and were analyzed with FlowJo software (Tree Star Inc., Ashland, Oregon, USA).