Ceacam1 Antibody / Carcinoembryonic antigen-related cell adhesion molecule 1 (FY13281)

Immunofluorescent staining of CEACAM1 using anti-Ceacam1 antibody (green). CEACAM1 was detected in an immunocytochemical section of mouse RAW264.7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-Ceacam1 antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. The section was counterstained with DAPI nuclear stain (blue). Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of CEACAM1 using anti-Ceacam1 antibody. Lane 1: rat liver tissue lysates, Lane 2: mouse Raw264.7 whole cell lysates, Lane 3: mouse Hepa1-6 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Ceacam1 antibody at 0.5 ug/ml overnight at 4oC, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal was developed using enhanced chemiluminescent. A predominant doublet is detected between an approximately 140 and 150 kDa in all samples, running well above the predicted ~57 kDa polypeptide mass. The higher apparent molecular weight is consistent with heavily N-glycosylated CEACAM1, and the 140-150 kDa doublet likely represents disulfide-linked dimers of mature CEACAM1 present in two closely spaced glycoforms.

Flow Cytometry analysis of HEPA1-6 cells using anti-Ceacam1 antibody. Overlay histogram showing HEPA1-6 cells stained with (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Ceacam1 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Flow Cytometry analysis of RAW264.7 cells using anti-Ceacam1 antibody. Overlay histogram showing RAW264.7 cells stained with (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-Ceacam1 antibody (1 ug/million cells) for 30 min at 20oC. DyLight 488 conjugated goat anti-rabbit IgG (5-10 ug/million cells) was used as secondary antibody for 30 minutes at 20oC. Isotype control antibody (Green line) was rabbit IgG (1 ug/million cells) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.