ERK1/ERK2 [p Thr185, p Tyr187, p Thr202, p Tyr204] Antibody [Unconjugated]

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.

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.

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.

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.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, Flow, IHC, CyTOF-ready
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

ERK1/ERK2 [p Thr185, p Tyr187, p Thr202, p Tyr204] Antibody [Unconjugated] Summary

• Description: Antibody detects ERK1 [p Thr202, p Tyr204] / ERK2 [p Thr185, p Tyr187]
• Immunogen: Phosphopeptide containing ERK1 T202/Y204 sites
• Modification: p Thr185, p Tyr187, p Thr202, p Tyr204
• Specificity: Detects human, mouse and rat Phospho-ERK1/ERK2 when dually phosphorylated at T202/Y204 and T185/Y187, respectively.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 0.1 ug/mL
  • Simple Western 5 ug/mL
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunohistochemistry 5-15 ug/mL
  • CyTOF-ready
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ERK1/ERK2 [p Thr185, p Tyr187, p Thr202, p Tyr204] Antibody [Unconjugated]

• ERK1/ERK2

Background

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.

1. Roskoski Jr., R. (2012) Pharmacol Res. 66:105.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ERK1/ERK2 Antibody (AF1018)(19)

Publications using AF1018

• JF Burgueño, JK Lang, AM Santander, I Fernández, E Fernández, J Zaias, MT Abreu Fluid supplementation accelerates epithelial repair during chemical colitis PLoS ONE, 2019;14(4):e0215387. 2019 [PMID: 31002683] (IHC-P, Mouse)
• SM Herbrich, S Kannan, RM Nolo, M Hornbaker, J Chandra, PA Zweidler-M Characterization of TRKA signaling in acute myeloid leukemia Oncotarget, 2018;9(53):30092-30105. 2018 [PMID: 30046390] (Western Blot, Human)
• Z Wan, C Xu, X Chen, H Xie, Z Li, J Wang, X Ji, H Chen, Q Ji, S Shaheen, Y Xu, F Wang, Z Tang, JS Zheng, W Chen, J Lou, W Liu PI(4,5)P2 determines the threshold of mechanical force-induced B cell activation J. Cell Biol., 2018;0(0):. 2018 [PMID: 29685902] (ICC, Mouse)
• NM Scheers, DIA Pereira, N Faria, JJ Powell Ferric citrate and ferric EDTA but not ferrous sulfate drive amphiregulin-mediated activation of the MAP kinase ERK in gut epithelial cancer cells Oncotarget, 2018;9(24):17066-17077. 2018 [PMID: 29682205] (Western Blot, Human)
• KM Wilson, AM Jagger, M Walker, E Seinkmane, JM Fox, R Kröger, P Genever, D Ungar Glycans modify mesenchymal stem cell differentiation to impact the function of resulting osteoblasts J. Cell. Sci., 2018;0(0):. 2018 [PMID: 29361539] (Western Blot, Human)
• BA Rodrigues, VR Muñoz, GK Kuga, RC Gaspar, SCBR Nakandakar, BM Crisol, JD Botezelli, LSS Pauli, ASR da Silva, LP de Moura, DE Cintra, ER Ropelle, JR Pauli Obesity Increases Mitogen-Activated Protein Kinase Phosphatase-3 Levels in the Hypothalamus of Mice Front Cell Neurosci, 2017;11(0):313. 2017 [PMID: 29062272] (Western Blot, Mouse)
• 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 Use of non-selective ?-blockers is associated with decreased tumor proliferative indices in early stage breast cancer Oncotarget, 2017;8(4):6446-6460. 2017 [PMID: 28031536] (Western Blot, Human)
• 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. 2016 [PMID: 26912655] (Western Blot, Human)
• Lutay N, Hakansson G, Alaridah N, Hallgren O, Westergren-Thorsson G, Godaly G Mycobacteria bypass mucosal NF-kB signalling to induce an epithelial anti-inflammatory IL-22 and IL-10 response. PLoS ONE, 2014;9(1):e86466. 2014 [PMID: 24489729] (Western Blot, Human)
• Nanjo, Shigeki, Yamada, Tadaaki, Nishihara, Hiroshi, Takeuchi, Shinji, Sano, Takako, Nakagawa, Takayuki, Ishikawa, Daisuke, Zhao, Lu, Ebi, Hiromich, Yasumoto, Kazuo, Matsumoto, Kunio, Yano, Seiji Ability of the Met kinase inhibitor crizotinib and new generation EGFR inhibitors to overcome resistance to EGFR inhibitors. PLoS ONE, 2013;8(12):e84700. 2013 [PMID: 24386407] (Western Blot, Human)
• Suenaga M, Yamamoto M, Tabata S, Itakura S, Miyata M, Hamasaki S, Furukawa T Influence of gefitinib and erlotinib on apoptosis and c-MYC expression in H23 lung cancer cells. Anticancer Res, 2013;33(4):1547-54. 2013 [PMID: 23564796] (Western Blot, Human)
• Clarke D, Al Ahmad A, Lee B, Parham C, Auckland L, Fertala A, Kahle M, Shaw C, Roberts J, Bix G Perlecan Domain V induces VEGF secretion in brain endothelial cells through integrin alpha5beta1 and ERK-dependent signaling pathways. PLoS ONE, 2012;7(9):e45257. 2012 [PMID: 23028886] (Western Blot, Human)
• Yasumoto K, Yamada T, Kawashima A, Wang W, Li Q, Donev IS, Tacheuchi S, Mouri H, Yamashita K, Ohtsubo K, Yano S The EGFR ligands amphiregulin and heparin-binding egf-like growth factor promote peritoneal carcinomatosis in CXCR4-expressing gastric cancer. Clin. Cancer Res., 2011;17(11):3619-30. 2011 [PMID: 21482691] (Western Blot, Human)
• Donev IS, Wang W, Yamada T, Li Q, Takeuchi S, Matsumoto K, Yamori T, Nishioka Y, Sone S, Yano S Transient PI3K Inhibition Induces Apoptosis and Overcomes HGF-Mediated Resistance to EGFR-TKIs in EGFR Mutant Lung Cancer. Clin. Cancer Res., 2011;17(8):2260-9. 2011 [PMID: 21220474] (Western Blot, Human)
• Jang SW, Liu X, Chan CB Deoxygedunin, a natural product with potent neurotrophic activity in mice. PLoS ONE, 2010;5(7):e11528. 2010 [PMID: 20644624] (Western Blot, Mouse)
• Popov D, Nemecz M, Dumitrescu M, Georgescu A, Bohmer FD Long-term high glucose concentration influences Akt, ERK1/2, and PTP1B protein expression in human aortic smooth muscle cells. Biochem. Biophys. Res. Commun., 2009;388(1):51-5. 2009 [PMID: 19647719] (Western Blot, Human)
• Montecucco F, Lenglet S, Braunersreuther V, Burger F, Pelli G, Bertolotto M, Mach F, Steffens S CB(2) cannabinoid receptor activation is cardioprotective in a mouse model of ischemia/reperfusion. J. Mol. Cell. Cardiol., 2009;46(5):612-20. 2009 [PMID: 19162037] (Western Blot, Mouse)
• 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 E-selectin regulates gene expression in metastatic colorectal carcinoma cells and enhances HMGB1 release. Int. J. Cancer, 2008;123(8):1741-50. 2008 [PMID: 18324679] (Western Blot, Human)
• Noser JA, Mael AA, Sakuma R, Ohmine S, Marcato P, Lee PW, Ikeda Y The RAS/Raf1/MEK/ERK signaling pathway facilitates VSV-mediated oncolysis: implication for the defective interferon response in cancer cells. Mol. Ther., 2007;15(8):1531-6. 2007 [PMID: 17505473] (Western Blot, Human)

Review for ERK1/ERK2 Antibody (AF1018) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Rat.

reviewed by: Anonymous WB Rat 01/05/2015

Summary

• Application: Western Blot
• Sample Tested: See PMID 23559668
• Species: Rat

Blocking

• Blocking Details: See PMID 23559668

Details

• Detection Notes: See PMID 23559668

Comments

• Comments: Published PMID 23559668