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Home/Search results/Products/ELISA Kits/Cell-Based ELISA Kits/Human/Mouse/Rat Phospho-ERK1/2 (T202/Y204) Cell-Based ELISA

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Description
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Antigen Information

Accession Number:

P27361
P28482

Gene ID:

5594
5595

Gene Symbols:

ERK1
ERK2
MAPK1
MAPK3
PRKM1
PRKM2
PRKM3

Protein Name & Synonyms:

Mitogen-activated protein kinase 3 (MAP kinase 3) (MAPK 3) (EC 2.7.11.24) (ERT2) (Extracellular signal-regulated kinase 1) (ERK-1) (Insulin-stimulated MAP2 kinase) (MAP kinase isoform p44) (p44-MAPK) (Microtubule-associated protein 2 kinase) (p44-ERK1)/Mitogen-activated protein kinase 1 (MAP kinase 1) (MAPK 1) (EC 2.7.11.24) (ERT1) (Extracellular signal-regulated kinase 2) (ERK-2) (MAP kinase isoform p42) (p42-MAPK) (Mitogen-activated protein kinase 2) (MAP kinase 2) (MAPK 2)

Assay Format

Pathway:

MAPK Signaling

Specificity:

The antibodies provided in this kit recognizes human, mouse and rat Erk1 phosphorylated at sites Thr202/Tyr204 and Erk2 phosphorylated at sites Thr185/Tyr187 and total Erk1/2 for comparison.

Number of Targets Detected:

Species Detected:

Human
Mouse
Rat

Compatible Sample Types:

Adherent Cell Culture

Design Principle:

Cell-based

Method of Detection:

Colorimetric

Quantitative/Semi-Quantitative:

Semi-Quantitative

Solid Support:

96-well Microplate

Product Specifications

Size:

1, 2, or 5 x 96-Well Microplate Kit

Product Features

Site and signal pathway-specific
In vitro detection of adherent cell culture
No sample lysis needed
Compatible with a standard ELISA plate reader
Faster results than with ELISA
Adaptable for high-throughput screening and drug discovery

Application Notes

Kit Components

Cell Culture Microplate
Wash Buffer A
Wash Buffer B
Fixing Solution
HRP-Conjugated Secondary Antibody
Blocking Buffer
TMB One-Step Substrate
Stop Solution
Quenching Buffer

Other Materials Required

Distilled or deionized water
100 ml and 1 liter graduated cylinders
Tubes to prepare sample dilutions
Protease and Phosphatase inhibitors
Precision pipettes to deliver 2 µl to 1 ml volumes
Adjustable 1-25 ml pipettes for reagent preparation
Benchtop rocker or shaker
Microplate reader capable of measuring absorbance at 450 nm

Protocol Outline

Seed 10,000-30,000 cells into each well and incubate overnight.
Apply various treatment, inhibitors or activators according to manufacture's instructions.
Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
Add 200 µl of Blocking Solution and incubate for 1 h at 37 °C.
Add 50 µl of 1X anti-phospho-protein specific antibody or anti-pan-protein specific antibody to each well and incubate for 2 h at RT.
Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
Add 100 µl of TMB One-Step Substrate Reagent to each well.
Incubate 30 min at RT.
Add 50 µl of Stop Solution to each well.
Read at 450 nm immediately.