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.

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

Target Name/s


Species Detected

Human, Mouse, Rat

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.


The antibody pair provided in this kit recognizes human, mouse and rat ERK1/2 (pThr202/pTyr204).



Antigen Information

Gene Symbol:
  • MAPK1

Product Specifications

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

Array/ELISA Format

  • MAPK Signaling
Species Detected:
  • Human
  • Mouse
  • Rat
Solid Support:
96-well Microplate
Design Principle:
  • Cell-based
Method of Detection:
Compatible Sample Types:
  • Adherent Cell Culture

  1. 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

  2. Chim H., Armijo BS., Miller E., et al. Propranolol induces regression of hemangioma cells through HIF-1α-mediated inhibition of VEGF-A. Ann Surg. 2012 Jul;256(1):146-56. doi: 10.1097/SLA.0b013e318254ce7a.
    Species: Human
    Sample Type: Cell Lysate

  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

  4. 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

  5. Guitton, Christophe, et al. "Protective cross talk between activated protein C and TNF signaling in vascular endothelial cells: implication of EPCR, noncanonical NF-κ, and ERK1/2 MAP kinases." American Journal of Physiology-Cell Physiology 300.4 (2011): C833-C842.

  6. 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

  7. 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

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

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