FOXC2 Antibody / Forkhead box protein C2 (FY12456)

Immunofluorescent staining of FOXC2 using anti-FOXC2 antibody (red) and anti-Tubulin Alpha antibody (green). FOXC2 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-FOXC2 antibody and mouse anti-Tubulin Alpha antibody overnight at 4oC. Cy3 Conjugated Goat Anti-Rabbit IgG and FITC 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 FOXC2 using anti-FOXC2 antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human Hela whole cell lysates, Lane 2: human 293T whole cell lysates, Lane 3: human whole cell lysates, Lane 4: human PC-3 whole cell lysates, Lane 5: rat PC-12 whole cell lysates, Lane 6: rat NRK whole cell lysates, Lane 7: mouse C2C12 whole cell lysates, Lane 8: mouse 3T3-L1 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-FOXC2 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 is developed using an ECL Plus Western Blotting Substratewith Tanon 5200 system. FOXC2 (~54 kDa predicted) was detected at ~58-60 kDa, consistent with phosphorylation- and SUMOylation-dependent mobility shifts reported for this transcription factor

Flow Cytometry analysis of Hela cells using anti-FOXC2 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-FOXC2 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.