Specificity and sensitivity of Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody. The antibody reacts specifically with as little as 0.25 ng of phosphorylated p42 MAP kinase and does not cross-react with up to 4 µg of nonphosphorylated p42 MAP kinase.
Western blot analysis of whole-cell extracts from unstarved wild-type mouse embryonic fibroblasts (MEFs) treated with the indicated combinations of basic Fibroblast Growth Factor (bFGF #9952, 100 ng/ml for 30 minutes), Platelet-Derived Growth Factor (PDGF #9909, 100 ng/ml for 30 minutes), MEK1 Inhibitor (PD98059 #9900, 50 µM, 2 hour pre-treatment), and MEK1/2 Inhibitor (U0126 #9903, 10 µM, 2 hour pre-treatment), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody #9101 (upper panel) and p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695 (lower panel).
Confocal immunofluorescent analysis of NIH/3T3 cells either U0126-treated (left) or PDGF-treated (right) and labeled with Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin #8953 (red).
Flow cytometric analysis of Jurkat cells, untreated (green) or PMA-treated (blue), using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) Antibody compared to a nonspecific negative control antibody (red).
Phosphorylated MEK and Erk were assayed in human peripheral blood lymphocytes stimulated with PMA in the presence or absence of the Raf inhibitor BAY 37-9751 or the MEK inhibitor U0126 #9903. BAY 37-951 blocked PMA-stimulated phosphorylation of both MEK and Erk, consistent with inhibition at the level of Raf, while U0126 blocked phosphorylation of Erk only, consistent with inhibition at the level of MEK. From Chow, S. et al. (2001) Cytometry 46, 72-78.