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.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
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.
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