Immunofluorescent analysis of Ntera-2 cells using CSB-MA134471A0m at a dilution of 1:100 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).

Overlay histogram showing Hela cells stained with CSB-MA134471A0m (red line). The cells were fixed with 70% Ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (10-¦g/1*10⁶cells) for 1 h at 4-¦C. The secondary antibody used was FITC goat anti-mouse IgG(H+L) at 1/200 dilution for 1 h at 4-¦C. Isotype control antibody (green line) was mouse IGg2b (10-¦g/1*10⁶cells) used under the same conditions. Acquisition of >10,000 events was performed.

Immunoprecipitating PODXL in HEK293 whole cell lysate
Lane 1: Mouse control IgG (1-¦g) instead of CSB-MA134471A0m in HEK293 whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: CSB-MA134471A0m (8-¦g) + HEK293 whole cell lysate (500-¦g)
Lane 3: HEK293 whole cell lysate (10-¦g)

Western Blot
Positive WB detected in: A549 whole cell lysate
All lanes: PODXL antibody at 2.5-¦g/ml
Secondary
Goat polyclonal to Mouse IgG at 1/5000 dilution
Predicted band size: 59, 56 kDa
Observed band size: 59 kDa