VALIDATION IMAGES
4% Paraformaldehyde-fixed Daudi (H) cell; Antibody incubation with (CD86) monoclonal Antibody, unconjugated (bsm-43589R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
The Daudi (H) cells were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.) , followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green): Rabbit Anti-CD86 antibody (bsm-43589R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
The MJ (H) cells were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.) , followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green): Rabbit Anti-CD86 antibody (bsm-43589R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
4% Paraformaldehyde-fixed MJ (H) cell; Antibody incubation with (CD86) monoclonal Antibody, unconjugated (bsm-43589R) 1:100, 90 min at 37°C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
4% Paraformaldehyde-fixed THP-1 (treated with TPA (80 nM, 18 h), rested in fresh medium for 48 h, and then stimulated with LPS (5 μg/mL, 18 h)) (H) cell; Antibody incubation with (CD86) monoclonal Antibody, unconjugated (bsm-43589R) 1:100, 90 min at 37°C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
The THP-1 (treated with TPA (80 nM, 18 h), rested in fresh medium for 48 h, and then stimulated with LPS (5 μg/mL, 18 h)) (H) cells were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.) , followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green): Rabbit Anti-CD86 antibody (bsm-43589R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.