FACE™ ERK1/2

by Active Motif
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Description

Fast Activated Cell-based ELISA (FACE™) Kits provide a simple, sensitive method for detecting protein phosphorylation directly in the cell, without making extracts or performing electrophoresis and membrane blotting. These 96-well, high-throughput assays are available in both colorimetric and chemiluminescent formats for over 20 different targets (see list at right). For complete details, click the FACE™ Method tab below.

FACE ERK1/2 Kits provide 96 rxns each of 2 antibodies that enable you to monitor and compare the levels of both phosphorylated and total ERK1/2. The phospho-ERK antibody recognizes ERK1 when phosphorylated at Thr202 and Tyr204, as well as ERK2 when phosphorylated at Thr185 and Tyr187; the total-ERK antibody recognizes ERK1 and ERK2 proteins regardless of their phosphorylation states. Click the ERK1/2 Info tab below for data and more information.

 
Name Format Cat No. Price  
FACE™ ERK1/2 1 x 96 rxns 48140 $525 Buy Now
FACE™ ERK1/2 5 x 96 rxns 48640 $2,100 Buy Now
FACE™ ERK1/2 Chemi 1 x 96 rxns 48240 $580 Buy Now
FACE™ ERK1/2 Chemi 5 x 96 rxns 48740 $2,320 Buy Now
Measurement of phosphorylated ERK1/2 (T202/Y204 & T185/Y187) and total ERK1/2 using the FACE ERK1/2 Cell Based ELISA (In Cell Western) method.
 
Figure 1: Colorimetric measurement of phosphorylated ERK and total ERK.

Murine Macrophage 4/4 cells were cultured in 96-well plates and serum-starved for 16 hours. Cells were then stimulated with the indicated amounts of Phorbol 12-myristate 13-acetate (PMA) for 10 minutes and fixed. Total ERK and phospho ERK were each assayed in triplicate using the phospho-ERK and total-ERK antibodies included in the FACE ERK1/2 Kit. Data was plotted after correction for cell number (performed through use of Crystal Violet). Note that the induction treatment did not affect the overall level of total ERK.

Antibody Specificities

The phospho-ERK antibody is specific for phosphorylated ERK1 and ERK2 and was raised against a synthetic phospho-peptide corresponding to residues Thr202 and Tyr204 of human ERK1 and Thr185 and Tyr187 of human ERK2. This antibody recognizes both phosphorylated ERK1 and ERK2. It does not recognize unphosphorylated ERK1 or ERK2, nor does it recognize other phosphorylated proteins. The total-ERK antibody recognizes ERK1 and ERK2 regardless of the phosphorylation state.

ERK1/2 Overview

The extracellular signal-regulated kinases 1 and 2 (ERK1 and ERK2), also called p44 and p42 MAP kinases, are members of the Mitogen Activated Protein Kinase (MAPK) family of proteins found in all eukaryotes. Because the 44 kDa ERK1

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