Anti-Human CCR5 Recombinant Antibody (MC-4) (CAT#: TAB-1541CL)

Figure 1 MC-4 inhibits CCR5 endocytosis mediated by chemokines.

(A and B) CHO-K1 cells expressing CCR5 were preincubated with medium (■), 10 μg/ml MC-4 (●), or MC-4 F(ab) (∆) for 30 min on ice and then incubated with RANTES (A) or AOP-RANTES (B) for 30 min at 37°C. Surface expression of CCR5 was measured by flow cytometry with the antibody MC-4 or MC-4 F(ab) (15 μg/ml, on ice) and FITC-conjugated secondary antibodies. Fluorescence was normalized as 100% in the absence of chemokines and 0% for background fluorescence. All experiments were repeated at least three times with similar results.

Blanpain, C., Vanderwinden, J. M., Cihak, J., Wittamer, V., Le Poul, E., Issafras, H.,... & Parmentier, M. (2002). Multiple active states and oligomerization of CCR5 revealed by functional properties of monoclonal antibodies. Molecular biology of the cell, 13(2), 723-737.

Figure 2 293T cells expressing CCR5-luc and CCR5-YFP were incubated with coelenterazine, and the luminescence and the fluorescence signals were quantified before and after the addition of the indicated mAb.

The BRET ratio was defined as [(emission at 485 nm ± 20)/(emission at 530 nm ± 25)]-[(emission at 485 nm ± 20)/(emission at 530 nm ± 25)] for CCR5-luc expressed alone in the same experiments. Variation in BRET signal compared with baseline (addition of buffer alone) after 10 min of incubation with antibodies is displayed. All experiments were repeated at least twice with similar results. All points were performed in triplicates (error bars: SEM).

Blanpain, C., Vanderwinden, J. M., Cihak, J., Wittamer, V., Le Poul, E., Issafras, H.,... & Parmentier, M. (2002). Multiple active states and oligomerization of CCR5 revealed by functional properties of monoclonal antibodies. Molecular biology of the cell, 13(2), 723-737.