HTRF ERK P-T202/Y204 KIT

Phospho-ERK assay kits are designed for the robust quantification of ERK modulation, phosphorylated on Thr202/Tyr204, for GPCR and RTK targeted screening as a MAPK Pathway readout. The simple mix-and-read protocol eliminates all wash steps for faster analysis and high quality output.

Specifications

How it works

Phospho-ERK (Thr202/Tyr204) assay principle

The Phospho-ERK (Thr202/Tyr204) assay measures ERK when phosphorylated at Thr202/Tyr204. Contrary to Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis or transfer. The Phospho-ERK (Thr202/Tyr204) assay uses 2 labeled antibodies: one with a donor fluorophore, the other one with an acceptor. The first antibody is selected for its specific binding to the phosphorylated motif on the protein, the second for its ability to recognize the protein independent of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both labeled antibodies and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of phosphorylated protein present in the sample, and provides a means of assessing the proteins phosphorylation state under a no-wash assay format.

phospho-erk-thr202-tyr204-assay-principle

Phospho-ERK 2-plate assay protocol

The 2 plate protocol involves culturing cells in a 96-well plate before lysis then transferring lysates to a 384-well low volume detection plate before adding Phospho-ERK HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.

phospho-erk-thr202-tyr204-assay-principle

Phospho-ERK 1-plate assay protocol

Detection of Phosphorylated ERK with HTRF reagents can be performed in a single plate used for culturing, stimulation and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality.

Simplified pathway

MAPK/ERK cell signaling

The MAPK/ERK signaling cascade is activated by a wide variety of receptors involved in growth and differentiation. The core signal transduction cascade elicits regulation of numerous cellular processes including adhesion, migration, apoptosis, differentiation and metabolism. MEK1/2 catalyze the phosphorylation of ERK1/2 at Tyr204 and Thr202. In response, ERK phosphorylates hundreds of cytoplasmic and nuclear substrates. The wide complexity and diversity of MAPK signaling makes ERK a key regulator and major signaling node in biology.

Revvity, we regard "impossibility" as inspiration and "inability" as the driving force. Revvity offers health science solutions, cutting - edge technologies and professional services, with its business covering the entire end - to - end process of scientific research exploration, development, diagnosis, and treatment. Relying on years of in - depth cultivation in fields such as translational multi - omics technology, biomarker identification, imaging, disease prediction, screening, detection and diagnosis, and informatics, Revvity is using the power of technology to break through the boundaries of human potential.

In 2023, Revvity's turnover exceeded $2.7 billion. It has more than 11,000 employees worldwide and provides diverse services for customers such as pharmaceutical and biotechnology enterprises, diagnostic laboratories, scientific research institutions and government agencies. The company is a member of the S&P 500 Index, and its customers are spread across more than 190 countries and regions around the world.

文件名称

立即下载

Cat.No	Name	Size
64ERKPEG	HTRF ERK P-T202/Y204 KIT	500 PTS
64ERKPEH	HTRF ERK P-T202/Y204 KIT	10K PTS