TFCP2 Antibody / Transcription factor CP2 (FY12147)

Flow Cytometry analysis of Caco-2 cells using anti-TFCP2 antibody. Overlay histogram showing Caco-2 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-TFCP2 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 TFCP2 using anti-TFCP2 antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human Caco-2 whole cell lysates, Lane 2: human THP-1 whole cell lysates, Lane 3: human RT4 whole cell lysates, Lane 4: human whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: 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-TFCP2 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 an ECL Plus Western Blotting Substrate. The expected band size for TFCP2 is at 57 kDa. It is commonly observed at 57-65 kDa, possibly showing multiple bands reflecting different phosphorylation states. A slightly larger hyperphosphorylated form may be observed in rodent samples.