TCF20 Antibody / Transcription factor 20 (FY12029)

Immunofluorescent staining of TCF20 using anti-TCF20 antibody (green) and anti-Beta Tubulin antibody (red). TCF20 was detected in an immunocytochemical section of U2OS cells. 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-TCF20 antibody and mouse anti-Beta Tubulin antibody overnight at 4oC. DyLight 488 Conjugated Goat Anti-Rabbit IgG and Cy3 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 TCF20 using anti-TCF20 antibody. Electrophoresis was performed on a 8% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human SIHA 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-TCF20 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. Expected size of TCF20 ~212 kDa. Observed band at ~250 kDa in human SiHa cell lysate. Although the exact ~250 kDa size is not extensively documented in peer-reviewed Western blot literature, this size shift aligns with antibody supplier validation (observed ~250 kDa) and is consistent with post-translational modifications or isoform variation of this large transcriptional co-regulator.