Antibody was affinity purified using an epitope specific to Timeless immobilized on solid support. The epitope recognized by A300-960A maps to a region between residue 1120 and 1170 of human Timeless using the numbering given in entry NP_003911.1 (GeneID 8914). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
hTIM, protein timeless homolog, TIM, TIM1, timeless circadian clock 1, timeless homolog, Tof1 homolog
between 1120 and 1170
2 - 8°C
Detection of human Timeless by western blot of immunoprecipitates. Samples: Whole cell lysate (0.5 or 1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-Timeless antibody A300-960A (lot A300-960A- 2 ) used for IP at 6 µg per reaction. Timeless was also immunoprecipitated by rabbit anti-Timeless antibody A300-961A. For blotting immunoprecipitated Timeless, A300-960A was used at 0.4 µg/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.