Apc conjugated human igg1

The analysis of CD44 protein expression was performed as reported in [55 (link)]. Briefly, HUVEC cells were harvested at a number of 1 × 10⁶ pellets were incubated in the dark on ice for 30 min in 100 µL of PBS containing APC-conjugated CD44 anti-human antibody (mouse monoclonal; BD Pharmigen, Franklin Lakes, NJ, USA), APC-conjugated human IgG1 (BD Pharmigen, Franklin Lakes, NJ, USA) was used as scrambled. The cells were analyzed with Becton Dickinson FACScan flow cytometer using the Cells Quest program, version 6.0.

WT, PGS and ANXA1 KO MIA PaCa-2 cells were harvested at a number of 1 × 10⁶ and analyzed for FPR-1, FPR-2 and CD44 proteins as previously described19 (link). Briefly, pellets were incubated on ice for 1 hour in PBS containing a primary antibody against FPR-1 (rabbit polyclonal, clone H-230; 1:500, Santa Cruz Biotechnology) or a primary antibody against FPR-2 (mouse monoclonal, clone GM-1D6; 1:100, Genovac). Then, cells incubated on ice for 1 hour in 100 μl of PBS containing AlexaFluor 488 anti-rabbit (1:1000; Molecular Probes) or Alexa-Fluor 488 anti-mouse (1:1000; Molecular Probes). To evaluate CD44 expression, cells were incubated on ice for 30 minutes in 100 μl of PBS containing APC-conjugated CD44 anti-human antibody (mouse monoclonal, clone G44–26; BD Pharmigen), APC-conjugated human IgG1 (BD Pharmigen) was used as scrambled. The cells were analyzed with Becton Dickinson FACScan flow cytometer using the Cells Quest program.