Product name :EnzyFluo™ ERK Phosphorylation Assay Kit

Catalog Number :EERK-100

Quantity :100 tests

Price :381 Eur

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Supplier :Bioassays

For quantitative fluorescent immunoenzymatic assay of ERK1/2 phosphorylation status in cultured cells.
Method: FL360/450nm, 530/585nm.
Samples: cells.
Species: human, mouse, rat..
Procedure: 3 hrs (hands-on time).
Size: 100 tests.

DESCRIPTION
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. 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) is measured
using a double immunoenzymatic labeling procedure.
Principle of Cell-Based ERK ELISA
A. Culture &
treat cells
B. Add Ab1 C. Add Ab2 D. Add fluorogenic
substrates. Measure
fluorescence mouse anti-phosphoERK intensity.
anti-rabbit IgG-ALP anti-mouse IgG-HRP
Primary Ab1: rabbit anti-total ERK
Secondary Ab2:
ALP (360/450nm)
pERK ERK pERK ERK
Y Y Y
pERK ERK
YY Y
pERK ERK
YY Y
HRP (530/585nm)
KEY FEATURES
Safe. Non-radioactive assay.
Simple and convenient. Total and pERK can be measured in the same
sample.
APPLICATIONS
Determination of ERK phosphorylation status in whole cells.
Evalutation of effects of ligands or drugs on ERK phosphorylation.
Species tested: human, mouse, rat.
KIT CONTENTS
10× Wash Buffer: 25 mL Blocking Buffer: 25 mL
ALP Substrate: 6 mL HRP Substrate: 6 mL
ERK-Ab1: 10 μL pERK-Ab1 10 μL
HRP-Ab2: 10 μL ALP-Ab2 10 μL
Storage conditions: The kit is shipped on ice. Store all reagents at -20°C.
Shelf life of 6 months after receipt.
Precautions: reagents are for research use only. Normal precautions for
laboratory reagents should be exercised while using the reagents. Please
refer to Material Safety Data Sheet for detailed information.

MATERIALS REQUIRED BUT NOT PROVIDED
37% formaldehyde (Sigma, cat # F8775); 3% H2O2 (Sigma, cat # H1009);
black cell culture 96-well plate: available separately at BioAssay Systems
(cat# P96BCC) or at Sigma (CLS3603); plate sealers: available separately
at BioAssay Systems (cat# AB96SL) or at Sigma (cat# A5596); deionized

datasheet of product :EnzyFluo™ ERK Phosphorylation Assay Kit EnzyFluo™ ERK Phosphorylation Assay Kit

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