| Assay Method Information | |
| | Electrophoretic Mobility Shift Assay (EMSA) |
| Description: | His10-Stat3 was expressed in Sf9 cells from a baculovirus encoding the recombinant protein. A nuclear extract of the Sf9 cells was incubated with 32P-labeled high affinity c-fos sis inducible element (hSIE) either alone or in the presence of inhibitor. After 20 min of incubation, samples were electrophoresed on polyacrylamide gels. The gels were dried, exposed to a phosphorimager screen and scanned. IC50 values were derived from plots of spot intensity versus phosphopeptide concentration. The affinity of peptides to the SH2 domain is measured by the intensity of the radioactivity of the Stat3-DNA complex band in the electrophoresis gel. |
| Affinity data for this assay | |
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