Agonist-mediated activation of endogenous MAP kinases. Agonist-mediated activation of MAP kinases (ERK1/2) was detected by immunoblotting performed on stably transfected cells expressing the fusion protein (HEK293/β2AR-Gαs) or the wild type β2-adrenergic receptor (HEK293/β2AR). Cells were first incubated in the absence (0) or presence of 1 μM isoproterenol (ISO) for the indicated times, and then whole-cell extracts were subjected to SDS-PAGE. The immunoreactivity of the endogenous ERK1/2 following agonist-induced phosphorylation was detected by probing blots with Phospho-p44/42 MAP kinases rabbit monoclonal antibody. Samples were normalized for protein content by reprobing blots with a monoclonal antibody to alpha-tubulin.
Di Certo et al. BMC Cell Biology 2008 9:56 doi:10.1186/1471-2121-9-56