IF analysis of CD68 and CD45 using anti-CD68 antibody (M00602) and anti-CD45 antibody (PB9785)
CD68 and CD45 was detected in paraffin-embedded section of human tonsil tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL mouse anti-CD68 Antibody (M00602) and rabbit anti CD45 Antibody(PB9785) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) and Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The tissue section was developed using Cy3 Conjugated Avidin (BA1037). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of A431 cells using anti-CD45 antibody (PB9785).
Overlay histogram showing A431 cells stained with PB9785 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD45 Antibody (PB9785, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.