Table 1.: Cell-based ERK Activation HTS Assays

Name Assay Principle Detection Technology Provider
In-cell Western Using ERK1/2 primary antibodies and infrared-labeled secondary antibodies sandwich assay to detect infrared signal in fixed cells Infrared fluorescence LI-COR Biosciences
MSD ERK Activation Assay Using ERK1/2 antibody conjugated with an electrochemiluminescent compound and capture antibody for phosphorylated ERK1/2 coated in microplate wells for detection Electrochemiluminesence Meso-Scale Discovery
AlphaScreen SureFire™Cellular ERK Assay Using antibody-coated AlphaScreen beads to generate a fluorescent signal when in close proximity due to binding of phospho-ERK1/2 (Thr202/Tyr204) Fluorescence emission TGR BioSciences & Perkin Elmer
Cayman ERK/MAPK (Thr202/Tyr204) Cell-based Assay A sandwich assay using anti-Thr202/Tyr204 primary antibody with a DyLight conjugated secondary antibody Immunofluorescence staining Cayman
Phospho-ERK (Cellul'erk) Assay A sandwich assay format using 2 different specific monoclonal antibodies: the anti-phospho-ERK antibody labeled with d2 and the anti-ERK antibody labeled with Eu3+-cryptate. HTRF Cisbio
LanthaScreen® Activity Assay Terbium (Tb)-labeled antibody binds the dually phosphorylated (Thr185, Tyr187) ERK2-GFP fusion protein expressed in cells. This association allows energy transfer to occur between the excited state Tb fluorophore and GFP, leading to an increase in TR-FRET signal TR-FRET Life Technologies