ERK Phosphorylation Assay
The mitogen-activated protein kinase (MAPK/ERK) pathway plays a key role in cell proliferation, differentiation and migration. Stimulation by mitogens eventually leads to phosphorylation of ERK1 (T202/Y204) and ERK2 (T185/Y187). The MAPK/ERK cascade presents many interesting drug targets for the development of cancer therapies.Assay Genie's cell-based ELISA ERK Phosphorylation Assay measures dually phosphorylated ERK1/2 in whole cells and normalizes the signal to the total protein content. This simple and efficient assay eliminates the need for cell lysate preparation and can be used to study kinase signaling and the effects of kinase inhibitors on cells. In this assay, cells are grown in 96-well plates and treated with ligands or drugs. Cells are then fixed and permeabilized in the wells. ERK1/2 phosphorylation (pERK) using a fluorescent ELISA followed by total protein measurement in each well.
ApplicationsFor quantitative fluorescent immunoenzymatic assay of ERK1/2 phosphorylation status in cultured cells.
- Safe. Non-radioactive assay.
- Simple and convenient. Total and pERK can be measured in the same sample.
ERK Phosphorylation Assay Information:
|Kit Includes:||10x Wash Buffer: 25 mL Blocking Buffer: 25 mL Protein Stain: 6 mL HRP Substrate: 6 mL HRP-Ab2: 10 uL pERK-Ab1: 10 uL|
|Kit Requires:||37% formaldehyde; 3% H2O2; black cell culture 96-well plate; plate sealers; deionized or distilled water; pipetting devices; cell culture incubators; centrifuge tubes; fluorescence plate reader capable of reading at ex/em = 530/585 nm and at ex/em =360/450 nm.|
|Method of Detection:||FL360/450nm, 530/585nm|
|Samples:||Cell, tissue etc|
|Species:||Human, mouse, rat.|
|Protocol Length:||8 hrs|
|Storage:||Store all reagents at -20°C|
|Shelf Life:||6 months|