NCR3LG1 Antibody / B7-H6 / Natural cytotoxicity triggering receptor 3 ligand 1 (FY12389)

Immunohistochemical staining of NCR3LG1 using anti-NCR3LG1 antibody. NCR3LG1 was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 ug/ml rabbit anti-NCR3LG1 antibody overnight at 4oC. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37oC. The tissue section was developed using an HRP secondary and DAB substrate.

Western blot analysis of NCR3LG1 using anti-NCR3LG1 antibody. Lane 1: human Hela whole cell lysates, Lane 2: human 293T whole cell lysates, Lane 3: human HepG2 whole cell lysates, Lane 4: human HEL whole cell lysates, Lane 5: rat testis tissue lysates, Lane 6: mouse testis tissue 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-NCR3LG1 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. NCR3LG1 (B7-H6) was detected at ~90 kDa, above the ~51 kDa predicted size, consistent with its documented N-linked glycosylation and mature membrane-glycoprotein form.

Flow Cytometry analysis of HepG2 cells using anti-NCR3LG1 antibody. Overlay histogram showing HepG2 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-NCR3LG1 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.