| Assay Method Information | |
| | mTOR Biochemical Assay |
| Description: | mTOR Biochemical PotencySolution of mTOR enzymes (Millipore, Cat. No. 14-770) was prepared in 1× kinase buffer at 4-fold of the final concentration (final concentration: 6 nM) in the assay. 2.5 μl of kinase solution was added to each well of the 384-well assay plate, which contains 2.5 μl of compounds with serially diluted concentration. 2× substrate solution was prepared with ULight-4E-BP1 (Thr37/46) Peptide (PE, Cat. No. TRF0128-M) and ATP in 1× kinase reaction buffer at 2-fold of the final concentration of each reagent in the assay. 5 μl of substrate solution was added to each well of the assay plate to start reaction. The assay plate was incubated at room temperature for 30 minutes. Detection solution of kinase quench buffer (EDTA) and Eu-anti-phospho-4E-BP1 antibody (Thr37/46) (PE, Cat. No. TRF0216-M) were prepared at 2-fold the desired final concentrations of each reagent in Lance detection buffer. 10 μl of detection solution buffer was added to each well of the assay plate. The assay plate was equilibrated for 60 minutes at room temperature before read on a plate reader (Lance signal (665 nm) from Envision program). |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |