PITPNM1 Antibody / Phosphatidylinositol transfer protein membrane associated 1 (FY12014)

Flow Cytometry analysis of Raji cells using anti-PITPNM1 antibody. Overlay histogram showing Raji 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-PITPNM1 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.

Western blot analysis of PITPNM1 using anti-PITPNM1 antibody. Lane 1: rat brain tissue lysates, Lane 2: mouse brain 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-PITPNM1 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. Expected size for PITPNM1 ~135 kDa. Observed strong band at ~150 kDa and a lighter band at ~120 kDa in mouse/rat brain lysate. The higher band likely corresponds to full-length or modified PITPNM1, while the lighter band may reflect a splice variant or truncated form. Alternative isoforms of PITPNM1 and membrane-associated processing of this multi-domain protein have been reported in the literature.