Example of a typical p70S6K (pT389) and p70S6K (Total) cell lysate dilution series. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
Example of a typical p70S6K (pT389) recombinant protein standard curve. The proportion of total protein that is phosphorylated is unknown - data is indicative only. Background-subtracted data values (mean +/- SD) are graphed.
. Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of p70S6K (pT389) are normalized and plotted.
Cell line analysis for Total p70S6K from 100 µg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
Induction of p70S6K (pT389) phosphorylation in MCF-7 cells in response to insulin treatment. MCF-7 cells were cultured in 96-well tissue culture plates, serum-starved and treated (10 min) with a dose-range of insulin before cell lysis. Data from quadruplicate measurements of p70S6K (pT389) are plotted and compared against Total p70S6K protein levels. Comparative p70S6K (pT389) and p70S6K (Total) data also shown by Western Blot.