discussion for serial dilution experiment

Cultures are usually allowed to grow through a normal growth curve, with daily transfer of a small volume of the expanded culture into fresh medium. Serial dilution generally refers to selection preformed in the standard growth regimes typically used in the lab: flasks, test tubes, solid media, or 96-well plates. Dunham, in Methods in Enzymology, 2010 3.1 Serial dilution The individual dissociation constant ( K d), if applicable, was derived from the T0901317 (10 μM) group by using the one-site total binding equation in the GraphPad Prism. The collected data (10,000 × 520 nm/490 nm) were fitted into a one-site total binding equation for the DMSO vehicle group, 10 μM T0901317 group, and DMSO vehicle group (without GST-PXR-LBD as the background control group) ( Fig. The TR-FRET signals of individual wells after 30, 60, 90, 120, and 180 min of incubation were measured with a PHERAstar FS plate reader. All assays were performed in the assay buffer (50 mM TRIS, pH 7.5, 20 mM MgCl 2, 0.1 mg/mL BSA, and 0.05 mM DTT). As an additional background control group, the same serial dilutions of FAM-SRC1-A, FAM-SRC1-B, and FAM-SRC1-C were incubated with Tb-anti-GST (5 nM), but with 1.1% DMSO (vehicle) and without GST-hPXR-LBD. The final DMSO concentration was 1.1% for all wells. Serial dilutions of FAM-SRC1-A, FAM-SRC1-B, and FAM-SRC1-C (1:2 dilutions giving 13 concentration levels from 2500 to 0.6 nM) were incubated with Tb-anti-GST (5 nM), GST-hPXR-LBD (5 nM), and either DMSO or T0901317 (10 μM). Wenwei Lin, Taosheng Chen, in Advances in Protein Chemistry and Structural Biology, 2018 3.1 PXR TR-FRET-Binding K d Determinations for the Fluorescent SRC1 Peptides FAM-SRC1-A, FAM-SRC1-B, and FAM-SRC1-C

Discussion for serial dilution experiment