Mouse anti cd43 antibody 1g10

HEK293T cells were transfected with 400 ng of pCMV3-PSGL-1, pCMV3-CD43, pCMV3-CD164, pCMV3-CD34, pCMV3-PODXL1, pCMV3-PODXL2, pCMV3-TIM-1, or pCMV3-Empty DNA. At 48 h post transfection, 0.5–1 million cells were stained with one of the following primary antibodies: mouse anti-PSGL1 antibody (KPL-1) (BD Pharmingen), mouse anti-CD43 antibody (1G10) (BD Biosciences), mouse anti-CD164 antibody (67D2) (Biolegend), mouse anti-CD34 antibody (563) (BD Biosciences), mouse anti-TIM-1 antibody (219211) (R & D Systems), mouse monoclonal anti-PODXL1 antibody (222328) (R & D systems), or mouse monoclonal anti-PODXL2 antibody (211816) (R & D Systems), followed by staining with Alexa Fluor 488-conjugated goat anti-mouse secondary antibody (Invitrogen).

HIV-1 particles were assembled in 6-well plates by co-transfection of HEK293T cells with pHIV-1(NL4-3) DNA (1 μg) plus an empty vector or the vector expressing each individual SHREK protein (400 ng). For MUC1 and TMEM123, vectors expressing heamagglutinin-tagged MUC1 or TMEM123 were used. Particles were harvested at 48 h, normalized for HIV-1 p24, and subjected to immuno-magnetic capture as previously described [22 (link)]. Briefly, magnetic Dynabeads Pan Mouse IgG (Invitrogen) (1 × 10⁸ beads/0.25 mL) were conjugated with one of the following antibodies, mouse anti-PSGL-1 antibody (KPL-1) (BD Pharmingen), mouse anti-CD43 antibody (1G10) (BD Biosciences, San Jose, CA, USA), mouse anti-CD164 antibody (67D2) (Biolegend, San Diego, CA, USA), mouse anti-PODXL1 antibody (222328) (R & D Systems, Minneapolis, MN, USA), mouse anti-PODXL2 antibody (R & D Systems), mouse anti-CD34 antibody (563) (BD Biosciences) or mouse anti-HA tag (HA.C5) antibody (Abcam, Cambridge, UK) for 30 min at room temperature. After conjugation, antibody-conjugated beads were incubated with SHREK-bearing viral particles for 1 h at 37 °C. The complex was pulled down with a magnet, and washed with cold PBS for 5 times. Captured viral particles were eluted in 10% Triton x-100 PBS, diluted, and quantified by p24 ELISA.