Human/Mouse/Rat Phospho-ERK1/2 (T202/Y204) Cell-Based ELISA

RayBio® Cell-Based Human/Mouse/Rat ERK1/2 (Thr202/Tyr204) Phosphorylation ELISA Kit. Suitable for adherent whole cell lines.


Assay Format

  • MAPK Signaling
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:
  • Semi-Quantitative
Solid Support:
96-well Microplate

Product Specifications

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

Protein 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 (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 (ERT1) (Extracellular signal-regulated kinase 2) (ERK-2) (MAP kinase isoform p42) (p42-MAPK) (Mitogen-activated protein kinase 2) (MAP kinase 2) (MAPK 2)

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
  1. Seed 10,000-30,000 cells into each well and incubate overnight.
  2. Apply various treatment, inhibitors or activators according to manufacture's instructions.
  3. Add 100 µl of Fixing Solution into each well and incubate for 20 min at RT with shaking.
  4. Add 200 µl of prepared 1X Quenching Buffer and incubate 20 min at RT.
  5. Add 200 µl of Blocking Solution and incubate for 1 h at 37 °C.
  6. 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.
  7. Add 50 µl of prepared 1X HRP-Anti-Rabbit or Mouse IgG and incubate for 1 h at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.


  1. Chim H., Armijo BS., Miller E., et al. Propranolol induces regression of hemangioma cells through HIF-1?-mediated inhibition of VEGF-A. Ann Surg. 2012 Jul;256(1):146-56. doi: 10.1097/SLA.0b013e318254ce7a.
    Species: Human
    Sample type: Cell Lysate (Hemangioma endothelial cells treated with propranolol)
  2. Soga F., et al. Serotonin Activates Human Monocytes and Prevents Apoptosis. J Invest Dermatol. 2007 Aug;127(8):1947-55.
    Species: Human
    Sample type: Cell Lysate (Monocytes)
  3. Flamein F, Riffault L, Muselet-Charlier C, et al. Molecular and cellular characteristics of ABCA3 mutations associated with diffuse parenchymal lung diseases in children. Human Molecular Genetics 2012;21(4):765-775. doi:10.1093/hmg/ddr508.
    Species: Human
    Sample type: Cell Lysate (Adenocarcinomic basal epithelial cell)
  4. Poggi, Marjorie, et al. "Palmitoylation of TNF alpha is involved in the regulation of TNF receptor 1 signalling." Biochimica et Biophysica Acta (BBA)-Molecular Cell Research 1833.3 (2013): 602-612.
    Species: Human
    Sample type: Conditioned Media (Hela, 3T3L1, ADAM17-deficient (ADAM17zn/zn) and RAW264.7 cell lines)
  5. Brown J., et al. Visfatin regulates insulin secretion, insulin receptor signalling and mRNA expression of diabetes-related genes in mouse pancreatic b-cells. Journal of Molecular Endocrinology (2010) 44, 171-178
    Species: Mouse
    Sample type: Cell Lysate (Beta-TC6 cells, BTC)
  6. Guitton, Christophe, et al. "Protective cross talk between activated protein C and TNF signaling in vascular endothelial cells: implication of EPCR, noncanonical NF-?B, and ERK1/2 MAP kinases." American Journal of Physiology-Cell Physiology 300.4 (2011): C833-C842.
    Species: Human
    Sample type: Cell Lysate
  7. Sharma U., et al. A novel role of alkaline phosphatase in the ERK1/2 dephosphorylation in renal cell carcinoma cell lines: A new plausible therapeutic target. Biochimie Sept 2014, epub ahead of print.
    Species: Human
    Sample type: Cell Lysate (Renal carcinoma cells)
  8. Menschikowski, Mario, et al. "Expression and shedding of endothelial protein C receptor in prostate cancer cells." Cancer cell international 11.1 (2011): 4.
    Species: Human
    Sample type: Conditioned Media
  9. Lerin R. Luckett-Chastain, Michael A. Ihnat, Bethany M. Mickle-Kawar, and Randle Michael Gallucci. SOCS3 Modulates Interleukin-6R Signaling Preference in Dermal Fibroblasts. J Interferon Cytokine Res. 2012 May;32(5):207-15. doi: 10.1089/jir.2011.0086
    Species: Mouse
    Sample type: Cell Lysate (Mouse pups fibroblast cells.)

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This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.