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

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    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.

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    Product Description

    Specifications

    Size 2 Plate Kit
    Species Human, Mouse, Rat
    Accession Number P27361|P28482|P45983|Q16539
    Gene Symbols MAPK3|ERK1|ERK2|PRKM1|PRKM3/MAPK1|PRKM2/MAPK8|JNK1|PRKM8|SAPK1|SAPK1C/MAPK14|CSBP|CSBP1|CSBP2|CSPB1|MXI2|SAPK2A
    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)
    Quantitative/Semi-Quantitative Semi-Quantitative
    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.
    Compatible Sample Types Adherent Cell Culture
    Solid Support 96-well Microplate
    Method Of Detection Colorimetric
    Design Principle Cell-based
    Research Area Post-Translational Modifications, Phosphorylation, MAPK Signaling
    Estimated Lead Time 1-2 business days
    Shipping Type Blue ice
    Storage -20°C

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    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

    Citations

    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 (Mouse WT or NLRP12-/- Dendritic cells)
    View Publication
    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 (Human Colon Cancer cell line, CCD18-Co)
    View Publication
    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 (Endothelial cells were stimulated with dsDNA )
    View Publication
    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 (RAW 264 cells, treated with DMTU)
    View Publication
    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 (H9c2 cells, rat ventricular myocardial cells.)
    View Publication
    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 (HepG2 Cells)
    View Publication
    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 (Synovial Cell, SW982)
    View Publication
    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 (Organ of corti (ear structure) lysates in a hearing loss model with drugs)
    View Publication

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