EnzyFluo™ ERK Phosphorylation Assay Kit

Protocol SDS

Application

• For quantitative fluorescent immunoenzymatic assay of ERK1/2 phosphorylation status in cultured cells.

Key Features

• New and improved. Total assay time reduced from the standard 21 hours to 6.5 hours (hands-on time 2.5 hrs).
• Simple and convenient. Cells are directly cultured in 96-well plates. No cell lysis is necessary.
• Accurate and high-throughput. Protein phosphorylation is normalized to total cellular protein in the same well, greatly minimizing well-to-well variations. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.

Method

• Cell-Based ELISA (FL360/450nm, 530/585nm)

Samples

• Cells

Species

• Human, mouse, rat

Procedure

• Assay takes 6.5 hrs, hands-on time 2.5 hrs

Size

• 100 tests

Shelf Life

• 6 months

More Details

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. BioAssay Systems cell-based ELISA 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.

Haven’t Found the ELISA Kit for Your Target

We offer two DIY kits: EFC1M-100 for mouse antibody and EFC2R-100 for rabbit antibody. All that is required is a primary antibody. If you prefer, we can develop the assay for you. Please email or call 1-510-782-9988 x 2 to request assay service.

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For more detailed product information and questions, please feel free to email us. Or for more general information regarding our assays, please refer to Technical Support.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.