CD300A Antibody (FY12806)

Flow Cytometry analysis of U20S cells using anti-CD300A antibody. Overlay histogram showing U20S 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-CD300A 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 CD300A using anti-CD300A antibody. Lane 1: human HL-60 whole cell lysates, Lane 2: human Raji whole cell lysates, Lane 3: rat PC-12 whole cell lysates, Lane 4: rat C6 whole cell lysates, Lane 5: mouse Raw264.7 whole cell lysates, Lane 6: mouse J774A.1 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-CD300A 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. Although the predicted molecular weight is ~33 kDa, CD300A is known to be heavily glycosylated and migrates at ~60-65 kDa under reducing conditions.