NKD1 Antibody / Naked cuticle 1 (FY12983)

Flow Cytometry analysis of Caco-2 cells using anti-NKD1 antibody. Overlay histogram showing Caco-2 cells stained with (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-NKD1 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.

Western blot analysis of NKD1 using anti-NKD1 antibody. Lane 1: human HepG2 whole cell lysates, Lane 2: human SW620 whole cell lysates, Lane 3: human Caco-2 whole cell lysates, Lane 4: human U20S 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-NKD1 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 specific band was detected for NKD1 at approximately 52 kDa. The expected molecular weight of NKD1 is ~52 kDa.