Human/Mouse/Rat Phospho-ERK1/2, JNK, p38 MAPK Cell-Based ELISA

$710.00
CBEL-ERK-SK

RayBio® Cell-Based Human/Mouse/Rat ERK1/2 (Thr202/Tyr204), JNK (Thr183/Tyr185), p38 MAPK (Thr180/Tyr182) 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

Erk1, Erk2, JNK, p38 MAPK

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.

Specificity

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

Storage/Stability

-20°C

Antigen Information

Gene Symbol:
  • MAPK1
  • MAPK8

Product Specifications

Size:
Two 96-Well Microplate Kit

Array/ELISA Format

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

  1. Arai M., et al. Protective Effects of Fucoidan Against Interleukin-1β-induced Inflammation in SW982 Human Synovial Cells. The Showa University Journal of Medical Sciences Vol. 26 (2014) No. 1 p. 47-56. http://dx.doi.org/10.15369/sujms.26.47
    Species: Human
    Sample Type: Cell Lysate

  2. Chen HM., Baicalein, an active component of Scutellaria baicalensis Georgi, prevents lysophosphatidylcholine-induced cardiac injury by reducing reactive oxygen species production, calcium overload and apoptosis via MAPK pathways. BMC Complementary and Alternative Medicine 2014, 14:233 doi:10.1186/1472-6882-14-233
    Species: Rat
    Sample Type: Cell Lysate

  3. Matsui S., et al. alcitonin gene-related peptide regulates mitogen-activated protein kinase pathway to decrease transforming growth factor β1-induced hepatic plasminogen activator inhibitor-1 mRNA expression in HepG2 cells. Biosci Biotech and Biochem. Volume 78, Issue 5, 2014 DOI: 10.1080/09168451.2014.910096
    Species: Human
    Sample Type: Cell Lysate

  4. Kim S, Yamamoto K, Nakamura Y, et al. A Possible Mechanism of Cisplatin-Induced Tumor Necrosis Factor (TNF)-α Production in Murine Macrophages. Pharmacology & Pharmacy. 2013;4(2):146-151.
    Species: Mouse
    Sample Type: Cell Lysate

  5. Bas E., Van De Water T., Gupta C., et al. Efficacy of three drugs for protecting against gentamicin-induced hair cell and hearing losses. Br J Pharmacol. 2012 Jul;166(6):1888-904. doi: 10.1111/j.1476-5381.2012.01890.x.
    Species: Rat
    Sample Type: Tissue Lysate

  6. Allen I., Wilson J., Schneider M., et al. NLRP12 suppresses colon inflammation and tumorigenesis through the negative regulation of noncanonical NF-κ signaling. Immunity. 2012 May 25;36(5):742-54. doi: 10.1016/j.immuni.2012.03.012.
    Species: Mouse
    Sample Type: Cell Lysate

  7. Gonzalez-Sarrias A., et al. NF-κ-dependent anti-inflammatory activity of urolithins, gut microbiota ellagic acid-derived metabolites, in human colonic fibroblasts. British Journal of Nutrition (2010), 104, 503-512
    Species: Human
    Sample Type: Cell Lysate

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Usage

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

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