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

$717.10

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.

CBEL-ERK-SK
+

Antigen Information

Accession Number:
  • P27361
  • P28482
  • P45983
  • Q16539
Gene Symbols:
CSBP | CSBP1 | CSBP2 | CSPB1 | ERK1 | ERK2 | JNK1 | MAPK3 | MXI2 | PRKM1   View more
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)/Mitogen-activated protein kinase 8 (MAP kinase 8) (MAPK 8) (EC 2.7.11.24) (JNK-46) (Stress-activated protein kinase 1c) (SAPK1c) (Stress-activated protein kinase JNK1) (c-Jun N-terminal kinase 1/Mitogen-activated protein kinase 14 (MAP kinase 14) (MAPK 14) (EC 2.7.11.24) (Cytokine suppressive anti-inflammatory drug-binding protein) (CSAID-binding protein) (CSBP) (MAP kinase MXI2) (MAX-interacting protein 2) (Mitogen-activated protein kinase p38 alpha) (MAP kinase p38 alpha) (Stress-activated protein kinase 2a) (SAPK2a)

Assay Format

Pathway:
  • MAPK Signaling
Specificity:
The antibodies provided in this kit recognizes human, mouse and rat Erk1 phosphorylated at sites Thr202/Tyr204, Erk2 phosphorylated at sites Thr185/Tyr187, JNK phosphorylated at sites Thr183/Tyr185 and p38 phosphorylated at sites Thr180/Tyr182. This kit also recognizes total Erk1/2, total JNK and total p38 for comparison.
Number of Targets Detected:
1
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:
Two 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
  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.

Storage/Stability

-20°C
  1. Arai M., et al. Protective Effects of Fucoidan Against Interleukin-1?-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 ?1-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)-? 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-?B signaling. Immunity. 2012 May 25;36(5):742-54. doi: 10.1016/j.immuni.2012.03.012.
    Species: Mouse
    Sample type: Cell Lysate
  7. Patel, Suraj J., et al. "The inflammatory response to double stranded DNA in endothelial cells is mediated by NF?B and TNF?." PloS one 6.5 (2011): e19910.
    Species: Human
    Sample type: Conditioned Media
  8. Gonzalez-Sarrias A., et al. NF-kB-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.