NSMCE4A Antibody / Non-SMC element 4 homolog A (FY12884)

Immunofluorescent staining of NSMCE4A using anti-NSMCE4A antibody (red) and anti-Beta Tubulin antibody (green). NSMCE4A was detected in immunocytochemical section of cell. 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-NSMCE4A antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. Cy3 Conjugated Goat Anti-Rabbit IgG and DyLight 488 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of NSMCE4A using anti-NSMCE4A antibody. Lane 1: human 293T whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: human whole cell lysates, Lane 4: human Jurkat whole cell lysates, Lane 5: rat ovary tissue lysates, Lane 6: rat RH35 whole cell lysates, Lane 7: mouse ovary tissue lysates, Lane 8: mouse NIH/3T3 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-NSMCE4A 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. NSMCE4A western blot shows a predominant band at ~50 kDa with a faint lower species. Although the predicted molecular weight is ~44 kDa, NSMCE4A frequently migrates higher due to phosphorylation and other post-translational modifications, producing a characteristic doublet near 48-50 kDa.

Flow Cytometry analysis of JK cells using anti-NSMCE4A antibody. Overlay histogram showing JK 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-NSMCE4A 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 (Red line) was also used as a control.