Product name :EnzyFluo™ Direct Phospho-ERK Assay Kit

Catalog Number :ERKD-100

Quantity :100 tests

Price :381 Eur

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Supplier :Bioassays

For quantitative direct fluorescent immunoenzymatic assay of ERK1/2 phosphorylation status in cultured cells.
Method: FL360/450nm, 530/585nm.
Samples: cells.
Species: human, mouse, rat.
Procedure: 2 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 normalizes the signal to the total protein
content. The assay is a useful tool to study the MAP kinase signaling
pathway. 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 HRP-conjugated
antibody specific for phosphorylated ERK1/2.
Principle of Direct pERK ELISA
A. Culture &
treat cells
pERK ERK
B. Add pERK-HRP
pERK ERK
C. Add HRP
substrate.
pERK ERK
HRP (530/585nm)
anti-pERK1/pERK-HRP: 2-HRP Conjugate HRP substrate: Protein Stain:
D. Add protein stain
E. Read fluorescence
Protein (360/450nm)
pERK ERK
Y
KEY FEATURES
Safe. Non-radioactive assay.
Simple and convenient. Saves time, direct one-step reaction, no secondary
antibody.
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
Protein Stain 6 mL HRP Substrate: 6 mL
pERK-HRP: 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.

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 System
(cat# P96BCC) or at Sigma (CLS3603); plate sealers: available separately
at BioAssay Systems (cat# AB96SL) or at Sigma (cat# A5596); deionized
or distilled water; pipetting devices; cell culture incubators; centrifuge tubes;
fluorescence plate reader capable of reading at lex/em = 530/585 nm and at
lex/em =360/450 nm.

datasheet of product :EnzyFluo™ Direct Phospho-ERK Assay Kit EnzyFluo™ Direct Phospho-ERK Assay Kit

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