Detection of Human E2F‑2 by Western Blot. |
Western blot shows lysates of HEK293 human embryonic kidney cell line. Gels were loaded with 30 μg of whole cell lysate (WCL), 20 μg of cytoplasmic (Cyto), and 10 μg of nuclear extracts (Nuc). PVDF membrane was probed with 1 µg/mL Goat Anti-Human E2F‑2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5119) followed by HRP‑conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band for E2F‑2 was detected at approximately 70 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
E2F-2 (viral E2-associated factor 2) is a 50‑55 kDa member of the E2F/DP family of transcription factors. It is principally expressed by placenta, and forms a DNA‑activating E2F heterodimeric complex with DP-1 or -2. This complex, when active, promotes cell cycle progression. In quiescent cells, association with the retinoblastoma-tumor suppressor gene product termed pRB suppresses its activity. Human E2F‑2 is 437 amino acids (aa) in length and contains a CDK2 binding region (aa 65‑105), a DNA‑binding domain (aa 107‑196), a dimerization segment (aa 197‑289), a transactivation region (aa 359‑437), and a pRB binding domain (aa 410‑427). There are two potential alternate start sites at Met197 and Met342, and one splice variant that shows a two aa substitution for aa 349‑437. Over aa 308‑437, human E2F-2 is 72% aa identical to mouse E2F-2.