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. 2007 Oct 29;28(1):511–527. doi: 10.1128/MCB.00800-07

FIG. 9.

FIG. 9.

Determination of HA-ERK1- and HA-ERK2-specific kinase activities. HA-ERK1 or HA-ERK2 was immunoprecipitated from increasing volumes of cell lysate obtained from CCL39 cells that stably express HA-ERK1 or HA-ERK2. An in vitro kinase assay using the GST-Elk-1307-428 fusion protein as the substrate was performed on each immunoprecipitate. (A) In the lysates of the kinase assays, the activated forms of HA-ERKs and the phosphorylation of the GST-Elk-1307-428 fusion protein were determined by immunoblotting using the anti-phospho-ERK1/2 and anti-phospho-Elk-1-ser383 antibodies, respectively. (B) The chemiluminescence was measured directly with a Gnome detector from Syngene (United Kingdom). The chemiluminescence corresponding to the phosphorylation of the GST-Elk-1307-428 fusion protein is plotted as a function of the chemiluminescence corresponding to the activated form of HA-ERK1 or HA-ERK2.