Antibody was affinity purified using an epitope specific to TSC1 immobilized on solid support. The epitope recognized by A300-316A maps to a region between residue 1100 and the C-terminus (residue 1164) of human tuberous sclerosis 1 using the numbering given in Swiss-Prot entry Q92574 (GeneID 7248). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
hamartin, LAM, truncated hemartin, TSC, tuberous sclerosis 1 protein
between 1100 and C-term
2 - 8°C
Detection of human and mouse TSC1 by western blot. Samples: Whole cell lysate (50 µg) from HeLa and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified goat anti-TSC1 antibody A300-316A (lot A300-316A-1) used for WB at 0.05 µg/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.
Detection of human and mouse TSC1 by western blot and immunoprecipitation. Samples: Whole cell lysate (10 or 50 µg for WB in A; 60 µg for input and 1.5 mg for IP in B) from HeLa (H), MEF (M), and U2OS cells. Antibody: Affinity purified rabbit anti-TSC1 antibody (Cat. No. A300-316A) used for WB at 0.2 (A & B) or 0.04 (A) µg/ml and used for IP at 1 µg/mg lysate. Detection: Chemiluminescence with an exposure time of 5 minutes (A) or 30 seconds (B). Note: TSC2 co-immunoprecipitates with TSC1. TSC2 was detected using a TSC2 antibody from another source.