Detection of Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) in Human PBMCs by Flow Cytometry.
Human peripheral blood mononuclear cells (PBMCs) either untreated (light orange filled histogram) or treated with 50 ng/mL PMA and 200 ng/mL Calcium Ionomycin for 10 minutes (dark orange filled histogram) were stained with Rabbit Anti-Human/Mouse/Rat Phospho-ERK1/ERK2 (ERK1 T202/Y204, ERK2 T185/Y187) Alexa Fluor® 488‑conjugated Monoclonal Antibody (Catalog # IC7806G) or isotype control antibody (Catalog # IC105G, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
ERK1 and ERK2 (also known as MAPK3 and MAPK1, respectively) are 44- and 42-kDa Ser/Thr kinases, respectively. They are part of the Ras-Raf-ERK signal transduction cascade often found downstream of growth factor receptor activation. ERK1 and ERK2 were initially isolated and cloned as kinases activated in response to Insulin and NGF. They are expressed in most, if not all, mammalian tissues. Dual threonine and tyrosine phosphorylation activate both ERKs, at Thr202/Tyr204 for human ERK1 and Thr185/Tyr187 for human ERK2. The two proteins share 83% amino acid identity, differing mainly at the N- and C-termini.
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