Example of a typical IĸBα (pS32/36) and IĸBα (Total) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.
Example of a typical IĸBα (pS32/36) and IĸBα (Total) 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 IĸBα (pS32/36) are normalized and plotted.
Cell line analysis for Total IĸBα from 150 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 IĸBα (pS32/36) phosphorylation in HeLa cells in response to TNFα treatment. HeLa cells were cultured in 96-well tissue culture plates, and treated (10 min) with a dose-range of TNFα before cell lysis. Data from quadruplicate measurements of IĸBα (pS32/36) are plotted and compared against Total IĸBα protein levels. Comparative IĸBα (pS32/36) and IĸBα (Total) data also shown by Western Blot.