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

(13 citations)   
  • Species Reactivity
    Human, Mouse, Rat
  • Specificity
    Detects human, mouse and rat Phospho-ERK1/ERK2 when dually phosphorylated at T202/Y204 and T185/Y187, respectively.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Phosphopeptide containing ERK1 T202/Y204 sites
  • 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
    0.1 µg/mL
    See below
  • Simple Western
    5 µg/mL
    See below
  • Flow Cytometry
    2.5 µg/106 cells
    See below
  • Immunohistochemistry
    5-15 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.
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 Human Phospho-ERK1 (T202/Y204) and ERK2 (T185/Y187) by Western Blot.

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 200 nM PMA for for the indicated times. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho‑ERK1 (T202/Y204)/ERK2 (T185/Y187) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1018), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho‑ERK1 (T202/Y204) and ERK2 (T185/Y187) at approximately 42 and 44 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Phospho-ERK1/ERK2 in Jurkat Human Cell Line by Flow Cytometry. Jurkat human acute T cell leukemia cell line were untreated (yellow line open histogram) or treated with 200 ng/mL PMA for 15 minutes (filled histogram) and stained with Rabbit Anti-Human/Mouse/Rat Phospho-ERK1/ERK2 (ERK1 T202/Y204, ERK2 T185/Y187) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1018) or control antibody (Catalog # AB‑105‑C, blue line open histogram), followed by Fluorescein-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0112). To facilitate intracellular staining, cells were fixed with PFA and permeabilized with ice-cold methanol.

Immunohistochemistry
ERK1/ERK2 in Rat Brain.

ERK1/ERK2 was detected in perfusion fixed frozen sections of rat brain (cortex) using 15 µg/mL Rabbit Anti-Human/Mouse/Rat Phospho-ERK1/ERK2 (ERK1 T202/Y204, ERK2 T185/Y187) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1018) overnight at 4 °C. Tissue was stained with the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Human Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) by Simple WesternTM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with PMA, loaded at 0.2 mg/mL. A specific band was detected for ERK1 (T202/Y204)/ERK2 (T185/Y187) at approximately 44 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse/Rat Phospho-ERK1 (T202/Y204)/ERK2 (T185/Y187) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1018). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 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 is a protein Serine/Threonine kinase that is a member of the extracellular signal-regulated kinases (ERKs) which are activated in response to numerous growth factors and cytokines (1). Activation of ERK1 requires both tyrosine and threonine phosphorylation that is mediated by MEK. ERK1 is ubiquitously distributed in tissues with the highest expression in heart, brain, and spinal cord. Activated ERK1 translocates into the nucleus where it phosphorylates various transcription factors.
  • References:
    1. Roskoski Jr., R. (2012) Pharmacol Res. 66:105.
  • 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.

13 Citations: Showing 1 - 10
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Species
Applications
Sample Type
  1. Use of non-selective ?-blockers is associated with decreased tumor proliferative indices in early stage breast cancer
    Authors: A Montoya, CN Amaya, A Belmont, N Diab, R Trevino, G Villanueva, S Rains, LA Sanchez, N Badri, S Otoukesh, A Khammanivo, D Liss, ST Baca, RJ Aguilera, EB Dickerson, A Torabi, AK Dwivedi, A Abbas, K Chambers, BA Bryan, Z Nahleh
    Oncotarget, 2017;8(4):6446-6460.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. Sirtuin 3 (SIRT3) Regulates ?-Smooth Muscle Actin (?-SMA) Production through the Succinate Dehydrogenase-G Protein-coupled Receptor 91 (GPR91) Pathway in Hepatic Stellate Cells
    J Biol Chem, 2016;291(19):10277-92.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Mycobacteria bypass mucosal NF-kB signalling to induce an epithelial anti-inflammatory IL-22 and IL-10 response.
    Authors: Lutay N, Hakansson G, Alaridah N, Hallgren O, Westergren-Thorsson G, Godaly G
    PLoS ONE, 2014;9(1):e86466.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  4. Ability of the Met kinase inhibitor crizotinib and new generation EGFR inhibitors to overcome resistance to EGFR inhibitors.
    Authors: Nanjo, Shigeki, Yamada, Tadaaki, Nishihara, Hiroshi, Takeuchi, Shinji, Sano, Takako, Nakagawa, Takayuki, Ishikawa, Daisuke, Zhao, Lu, Ebi, Hiromich, Yasumoto, Kazuo, Matsumoto, Kunio, Yano, Seiji
    PLoS ONE, 2013;8(12):e84700.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  5. Influence of gefitinib and erlotinib on apoptosis and c-MYC expression in H23 lung cancer cells.
    Authors: Suenaga M, Yamamoto M, Tabata S, Itakura S, Miyata M, Hamasaki S, Furukawa T
    Anticancer Res, 2013;33(4):1547-54.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  6. Perlecan Domain V induces VEGF secretion in brain endothelial cells through integrin alpha5beta1 and ERK-dependent signaling pathways.
    Authors: Clarke D, Al Ahmad A, Lee B, Parham C, Auckland L, Fertala A, Kahle M, Shaw C, Roberts J, Bix G
    PLoS ONE, 2012;7(9):e45257.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  7. Transient PI3K Inhibition Induces Apoptosis and Overcomes HGF-Mediated Resistance to EGFR-TKIs in EGFR Mutant Lung Cancer.
    Authors: Donev IS, Wang W, Yamada T, Li Q, Takeuchi S, Matsumoto K, Yamori T, Nishioka Y, Sone S, Yano S
    Clin. Cancer Res., 2011;17(8):2260-9.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  8. The EGFR ligands amphiregulin and heparin-binding egf-like growth factor promote peritoneal carcinomatosis in CXCR4-expressing gastric cancer.
    Authors: Yasumoto K, Yamada T, Kawashima A, Wang W, Li Q, Donev IS, Tacheuchi S, Mouri H, Yamashita K, Ohtsubo K, Yano S
    Clin. Cancer Res., 2011;17(11):3619-30.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  9. Deoxygedunin, a natural product with potent neurotrophic activity in mice.
    Authors: Jang SW, Liu X, Chan CB
    PLoS ONE, 2010;5(7):e11528.
    Species: Mouse
    Sample Type: Cell Lysates
    Application: WB
  10. Long-term high glucose concentration influences Akt, ERK1/2, and PTP1B protein expression in human aortic smooth muscle cells.
    Authors: Popov D, Nemecz M, Dumitrescu M, Georgescu A, Bohmer FD
    Biochem. Biophys. Res. Commun., 2009;388(1):51-5.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  11. CB(2) cannabinoid receptor activation is cardioprotective in a mouse model of ischemia/reperfusion.
    Authors: Montecucco F, Lenglet S, Braunersreuther V, Burger F, Pelli G, Bertolotto M, Mach F, Steffens S
    J. Mol. Cell. Cardiol., 2009;46(5):612-20.
    Species: Mouse
    Sample Type: Tissue Homogenates
    Application: WB
  12. E-selectin regulates gene expression in metastatic colorectal carcinoma cells and enhances HMGB1 release.
    Authors: Aychek T, Miller K, Sagi-Assif O, Levy-Nissenbaum O, Israeli-Amit M, Pasmanik-Chor M, Jacob-Hirsch J, Amariglio N, Rechavi G, Witz IP
    Int. J. Cancer, 2008;123(8):1741-50.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  13. The RAS/Raf1/MEK/ERK signaling pathway facilitates VSV-mediated oncolysis: implication for the defective interferon response in cancer cells.
    Authors: Noser JA, Mael AA, Sakuma R, Ohmine S, Marcato P, Lee PW, Ikeda Y
    Mol. Ther., 2007;15(8):1531-6.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
Expand to show all 13 Citations
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