FOXM1 Antibody / Forkhead box protein M1 (FY12211)

Immunofluorescent staining of FOXM1 using anti-FOXM1 antibody (green) and anti-Beta Tubulin antibody (red). FOXM1 was detected in immunocytochemical section of HELA cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 ug/ml rabbit anti-FOXM1 antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG and DyLight 550 Conjugated Goat Anti-Mouse IgG were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37oC. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Western blot analysis of FOXM1 using anti-FOXM1 antibody. Lane 1: human Hela whole cell lysates, Lane 2: human DLD-1 whole cell lysates, Lane 3: human MCF-7 whole cell lysates, Lane 4: human U2OS whole cell lysates, Lane 5: rat thymus tissue lysates, Lane 6: rat testis tissue lysates, Lane 7: mouse thymus tissue lysates, Lane 8: mouse testis 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-FOXM1 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. The expected band size for FOXM1 is at 85-100 kDa (multiple isoforms).

Flow Cytometry analysis of Hela cells using anti-FOXM1 antibody. Overlay histogram showing Hela 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-FOXM1 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.