Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antibody

(6 citations)
(1 Review)
  
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse, and rat ERK1 and ERK2 dually phosphorylated at T202/Y204 or T185/Y187, respectively.
  • Source
    Monoclonal Rabbit IgG Clone # 269434
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Phosphopeptide containing ERK1 T202/Y204 site
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Simple Western
    5 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    8-25 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Mouse, Rat, and Human Phospho-ERK1 (T202/Y204) and ERK2 (T185/Y187) by Western Blot. Western blot shows lysates of NIH‑3T3 mouse embryonic fibroblast cell line untreated (-) or treated (+) with 100 ng/mL Rabbit Anti-Human PDGF (Catalog # 120-HD) and PC‑12 rat adrenal pheochromocytoma cell line untreated or treated with 100 ng/mL Recombinant Rat beta -NGF (Catalog #
556‑NG) and HeLa human cervical epithelial carcinoma cell line untreated or treated with 200 nM PMA. PVDF membrane was probed with 0.5 µg/mL of Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Monoclonal Antibody (MAB1018), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho-ERK1 (T202/Y204)/ ERK2 (T185/Y187) at approximately 40-45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 3.
Immunocytochemistry
Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) in HeLa Human Cell Line. ERK1/ERK2 phosphorylated at T202/Y204 (ERK1) and T185/Y187 (ERK2) was detected in immersion fixed Hela human cervical epithelial carcinoma cells, unstimulated (lower panel) and stimulated (upper panel) with PMA using Rabbit Anti-Human/Mouse/Rat Phospho-ERK1/ERK2 (ERK1 T202/Y204, ERK2 T185/Y187) Monoclonal Antibody (Catalog # MAB1018) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Intracellular Staining by Flow Cytometry
Detection of Phospho-ERK1/ERK2 in PMA-treated Jurkat Human Cell Line by Flow Cytometry. Jurkat human acute T cell leukemia cell line were unstimulated (light orange open histogram) or treated with 50 ug/mL PMA for 10 minutes (dark orange filled histogram), then stained with Rabbit Anti-Human/Mouse/Rat Phospho-ERK1/ERK2 (ERK1 T202/Y204, ERK2 T185/Y187) Monoclonal Antibody (Catalog # MAB1018) or control antibody (Catalog # AB‑105‑C, blue open histogram), followed by Fluorescein-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0112). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with ice-cold methanol.
Detection of Human Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) by Simple WesternTM. Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line untreated (-) or treated (+) with 200 nM PMA and Ionomycin for 20 minutes, loaded at 0.2 mg/mL. A specific band was detected for Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) at approximately 42 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-ERK1/ERK2 (ERK1 T202/Y204, ERK2 T185/Y187) Monoclonal Antibody (Catalog # MAB1018). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: ERK1/ERK2
ERK1 and ERK2 (also known as MAPK3 and MAPK1) are 44- and 42-kDa Ser/Thr kinases, respectively. They are part of the Ras-Raf-ERK signal transduction cascade often found downstream of growth factor receptor activation. ERK1 and ERK2 were initially isolated and cloned as kinases activated in response to insulin and NGF. They are expressed in most, if not all, mammalian tissues. Dual threonine and tyrosine phosphorylation activate both ERKs, at Thr202/Tyr204 for human ERK1 and Thr185/Tyr187 for human ERK2. The two proteins share 83% amino acid identity, differing mainly at the N and C termini.
  • Long Name:
    Extracellular Signal-regulated Kinase 1/2
  • Alternate Names:
    ERK1/ERK2
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

6 Citations: Showing 1 - 6
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Species
Applications
Sample Type
  1. Beta 1-integrin-c-Met cooperation reveals an inside-in survival signalling on autophagy-related endomembranes
    Authors: Rachel Barrow-McG
    Nat Commun, 2016;7(0):11942.
  2. T cell Ig and mucin domain-containing protein 3 is recruited to the immune synapse, disrupts stable synapse formation, and associates with receptor phosphatases.
    Authors: Clayton K, Haaland M, Douglas-Vail M, Mujib S, Chew G, Ndhlovu L, Ostrowski M
    J Immunol, 2014;192(2):782-91.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC Fluorescence
  3. Annexin A10 in human oral cancer: biomarker for tumoral growth via G1/S transition by targeting MAPK signaling pathways.
    Authors: Shimizu T, Kasamatsu A, Yamamoto A, Koike K, Ishige S, Takatori H, Sakamoto Y, Ogawara K, Shiiba M, Tanzawa H, Uzawa K
    PLoS ONE, 2012;7(9):e45510.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. Cellular repressor of E1A-stimulated genes inhibits human vascular smooth muscle cell apoptosis via blocking P38/JNK MAP kinase activation.
    Authors: Han Y, Wu G, Deng J, Tao J, Guo L, Tian X, Kang J, Zhang X, Yan C
    J. Mol. Cell. Cardiol., 2010;48(6):1225-35.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  5. The myeloperoxidase-derived oxidant HOSCN inhibits protein tyrosine phosphatases and modulates cell signalling via the mitogen-activated protein kinase (MAPK) pathway in macrophages.
    Authors: Lane AE, Tan JT, Hawkins CL
    Biochem. J., 2010;430(1):161-9.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  6. The involvement of the fractalkine receptor in the transmigration of neuroblastoma cells through bone-marrow endothelial cells.
    Authors: Nevo I, Sagi-Assif O, Meshel T, Ben-Baruch A, Johrer K, Greil R, Trejo LE, Kharenko O, Feinmesser M, Yron I, Witz IP
    Cancer Lett., 2009;273(1):127-39.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
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Average Rating: 5 (Based on 1 review)

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We have 1 review tested in 1 application: Western Blot.

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Summary

ApplicationWestern Blot
Sample TestedCell Extracts
SpeciesHuman

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