Western blot analysis of CD86 on different lysates with Rabbit anti-CD86 antibody (HA750109) at 1/5,000 dilution. Lane 1: Raji cell lysate (10 µg/Lane) Lane 2: Mouse lung tissue lysate (20 µg/Lane) Lane 3: Rat lung tissue lysate (20 µg/Lane) Predicted band size: 38 kDa Observed band size: 70 kDa Exposure time: 1 minute; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750109) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

Immunocytochemistry analysis of Raji cells labeling CD86 with Rabbit anti-CD86 antibody (HA750109) at 1/500 dilution. Cells were fixed in 80% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD86 antibody (HA750109) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.