Human U937 Cells were fixed with 2% PFA (10min at room temperature). The cells were then incubated in 1X PBS containing 0.5% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min at room temperature. Staining was performed with CD86 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:100 for 30 min on ice. A Goat Anti-rabbit IgG-PE secondary antibody was used for a 30 min incubation on ice. Primary antibody staining (green) is compared to unstained cells (dark blue), secondary antibody only (light blue) and isotype control (bs-0295P; orange).


Mouse Brain cells probed with CD86 Polyclonal Antibody, PE Conjugated (bs-1035R-PE; green) at 1:50 compared to isotype control (bs-0295P-PE; orange).


L1 rat brain lysates L2 rat heart lysates probed with Anti CD86/B7-2 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:200 overnight at 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 for 90 min at RT. Predicted band 34kD. Observed band size:34kD.


Raji cell lysates probed with CD86 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:300 overnight at 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:500 for 90 min at 37˚C.


Mouse BV-2 Cells were fixed with 4% PFA (10min at room temperature) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. The cells were then incubated in 1X PBS / 2% BSA / 10% goat serum to block non-specific protein-protein interactions for 20 min at 37°C. The cells were stained with CD86 Polyclonal Antibody Unconjugated (bs-1035R; green) at 1:100 for 90 min at 37°C. A FITC-conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) was incubated at 37°C for 90 minutes. DAPI (5ug/ml; blue) was used to stain the cell nuclei.


Lane 1: Human HepG2 cell lysates; Lane 2: Human U937 cell lysates; Lane 3: Rat Spleen lysates; Lane 4: Mouse Spleen lysates; Lane 5: Human HL-60 cell lysates probed with CD86 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.


The Daudi(H) cells were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Primary Antibody (green): Rabbit Anti-CD86 antibody (bs-1035R): 1 μg/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-60295G-FITC): 1 μg/test. 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 (bs-1035R): 1 μg/10^6 cells; Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.


4% Paraformaldehyde-fixed Daudi (H) cell; Antibody incubation with (CD86) polyclonal Antibody, unconjugated (bs-1035R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-40295G-FITC) 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.


25 ug total protein per lane of various lysates (see on figure) probed with CD86 polyclonal antibody, unconjugated (bs-1035R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.