Tuberous sclerosis 1 (TSC1), also known as hamartin, is a protein that in humans is encoded by the TSC1 gene. TSC1 functions as a co-chaperone which inhibits the ATPase activity of the chaperone Hsp90 (heat shock protein-90) and decelerates its chaperone cycle. TSC1 functions as a facilitator of Hsp90 in chaperoning the kinase and non-kinase clients including TSC2, therefore preventing their ubiquitination and degradation in the proteasome. TSC1, TSC2 and TBC1D7 is a multi-protein complex also known as the TSC complex. This complex negatively regulates mTORC1 signaling by functioning as a GTPase-activating protein (GAP) for the small GTPase Rheb, an essential activator of mTORC1. The TSC complex has been implicated as a tumor suppressor.
Background References
1. Li S et al. Hamartin: An Endogenous Neuroprotective Molecule Induced by Hypoxic Preconditioning. Front Genet. 2020 Sep
Western blot analysis of Hamartin on different lysates with Rabbit anti-Hamartin antibody (HA723665) at 1/20,000 dilution.
Lane 1: HeLa cell lysate Lane 2: 293T cell lysate Lane 3: COS-1 cell lysate Lane 4: NIH/3T3 cell lysate Lane 5: MEF cell lysate Lane 6: C2C12 cell lysate Lane 7: PC-12 cell lysate Lane 8: C6 cell lysate Lane 9: Mouse brain tissue lysate Lane 10: Mouse heart tissue lysate Lane 11: Rat brain tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 130 kDa Observed band size: 140 kDa
Exposure time: 1 minute; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723665) at 1/20,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Hamartin was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with HA723665 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723665 at 1/20,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: NIH/3T3 cell lysate (input) Lane 2: HA723665 IP in NIH/3T3 cell lysate Lane 3: Rabbit IgG instead of HA723665 in NIH/3T3 cell lysate