Detection of ERK5, phosphorylated on Thr218 and Tyr 220, by immunoblotting. Samples: Whole cell lysate from HEK293 cells transiently transfected with plasmids expressing the ERK5 kinase domain and constitutively activated MEK5D-D. Primary antibody: PhosphoDetect™ Anti-ERK5 (Thr218/Tyr 220) Rabbit pAb (Cat. No. 442688) (1:000) pre-incubated without peptide (lane 1), with non-phosphopeptide corresponding to the immunogen (lane 2), a generic phosphothereonine-containing peptide (lane 3), a generic phosphotyrosine-containing peptide (lane 4), phosphopeptide derived from the corresponding region of ERK1&2 (lane 5), or phosphopeptide immunogen (lane 6). Detection: chemiluminescence.
|
