|Detection of Human TSC2 by Western Blot. Western blot shows lysates of 293T human embryonic kidney cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human TSC2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4040) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for TSC2 at approximately 200 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|TSC2 in Human Kidney. TSC2 was detected in immersion fixed paraffin-embedded sections of human kidney using Sheep Anti-Human TSC2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4040) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
TSC-2 (Tuberous sclerosis complex protein 2; also tuberin) is a 190-220 kDa intracellular GTPase activating protein. It is widely expressed, and plays a central role in mTOR activation. In quiescent cells, TSC-2 is non-covalently bound to both membrane-embedded TSC-1 (a structurally unrelated protein) and polycystin-1. In this complex, TSC-2 represses mTOR activity by converting GTP-Rheb to GDP-Rheb. When AKT is activated (via growth factor signaling), TSC-2 is phosphorylated, binds to 14-3-3 protein, dissociates from its complex, and stops converting GTP-Rheb to GDP-Rheb. Human TSC-2 is 1807 amino acids (aa) in length. It contains two coiled-coil regions (aa 346-371 and 1008-1021), and one Rap-GAP domain (aa 1531-1758). Phosphorylation regulates TSC-2 activity and there are at least 19 utilized phosphorylation sites. Multiple splice variants exist. Singularly, or in combination, there are deletions of aa 1272-1294, 76-112, 946-988, and 946-989. Single point mutations abound, some of which impact phosphorylation. There is also a potential alternative start site 91 aa upstream of the standard site. Over aa 1506-1748, human TSC-2 shares 85% aa identity with mouse TSC-2.