Immunohistochemical analysis of paraffin-embedded human liver tissue using BD-PB2383 performed on the Leica® BOND RXm. Tissue was fixed with formaldehyde at room temperature; antigen retrieval was by heat mediation with a EDTA buffer (pH9. 0). Samples were incubated with primary antibody(1:1000) for 1 hours at room temperature. A undiluted biotinylated CRF Anti-Polyvalent HRP Polymer antibody was used as the secondary antibody.

Overlay histogram showing Jurkat cells stained with BD-PB2383(green line). The cells were fixed with 2% paraformaldehyde and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed at 1/200 dilution for 40 min at Room temperature. Isotype control antibody (blue line) was rabbit IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

All lanes : Anti-CD14 Antibody (N-term) at 1:2000 dilution Lane 1: Human liver lysate Lane 2: Human lung lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 40 kDa Blocking/Dilution buffer: 5% NFDM/TBST.