LSP1 Antibody / Lymphocyte-specific protein 1 (FY12761)

Flow Cytometry analysis of U937 cells using anti-LSP1 antibody. Overlay histogram showing U937 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-LSP1 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 LSP1 using anti-LSP1 antibody. Lane 1: human HL-60 whole cell lysates, Lane 2: human U-937 whole cell lysates, Lane 3: human Raji whole cell lysates, Lane 4: human Daudi whole cell lysates, Lane 5: rat spleen tissue lysates, Lane 6: rat thymus tissue lysates, Lane 7: mouse spleen tissue lysates, Lane 8: mouse thymus 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-LSP1 antibody at 0.25 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 band is detected at ~50 kDa, consistent with the phosphorylated form of LSP1 (predicted molecular weight ~37 kDa). The apparent upward shift is characteristic of LSP1's phosphorylation-dependent migration behavior observed in immune cell lysates.