CD14 Antibody (N-term)

Immunohistochemical analysis of paraffin-embedded human liver tissue was 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.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 cells labeling CD14 at 1/25 dilution, followed by Dylight 488-conjugated goat anti-Rabbit IgG secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm and membrance staining on A549 cell line. Cytoplasmic actin is detected with Dylight 554 Phalloidin (1186255) at 1/500 dilution (red). The nuclear counter stain is DAPI (blue).

Overlay histogram showing Jurkat cells stained (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 37°C. 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 (1 μg/1x10^6 cells) used under the same conditions. Acquisition of > 10000 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.

All lanes: Anti-CD14 Antibody (N-term) at 1:1000 dilution. Lane 1: Human lung tissue lysate. Lane 2: Human liver tissue 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.

All lanes: Anti-CD14 Antibody (N-term) at 1:2000 dilution. Lane 1: Human lung tissue lysate. Lane 2: Human liver tissue lysate. Lane 3: A549 whole cell 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.

All lanes: Anti-CD14 Antibody (N-term) at 1:1000 dilution. Lane 1: THP-1 whole cell lysate. Lane 2: Human lung tissue lysate. Lane 3: Human liver tissue lysate. Lane 4: A549 whole cell 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.