详细信息:Millipore’s FlowCellect™ EGFR/MAPK Activation Detection kit includes two directly conjugated phospho specific antibodies: pEGFR(Tyr1173)-Alexa Fluor 488 and pERK (Thr202/Tyr204, Thr185/Tyr187)-PE, along with optimized fixation, permeabilization, wash and assay buffers to provide researchers the ability to detect the activation of EGFR and ERK at the receptor and downstream signaling level.
The ability to detect the phosphorylation of EGFR and ERK simultaneously allows researchers to profile EGF mediated MAPK activation within a cell type of interest. Alterations in these signaling profiles can be used to monitor the effect of a particular upstream treatment targeting the pathway.
Antibodies against phospho-EGFR and phospho-ERK1/2 are provided in the kit to measure the extent of each proteins activation state. A431 cells were used as a model system to demonstrate EGF mediated activation of EGFR as well as its’ downstream effector kinase ERK. The treated cells were compared to untreated cells to verify the detection of both activated proteins by flow cytometry. When performing a dual color stain, the majority of the treated cells are double positive indicating activation of MAPK through EGFR.
Sufficient reagents are provided for 25 3-color samples. The kit includes all optimized fluorescently labeled antibodies and buffers necessary for cell preparation and analysis.
