ERK1/ERK2 [p Tyr187, p Tyr204, p Thr185, p Thr202] Antibody (269434) [Unconjugated]

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.

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.

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.

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.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, CyTOF-ready, Flow, ICC/IF
• Clone: 269434
• Clonality: Monoclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

ERK1/ERK2 [p Tyr187, p Tyr204, p Thr185, p Thr202] Antibody (269434) [Unconjugated] Summary

• Description: Antibody detects ERK1 [p Thr202, p Tyr204] / ERK2 [p Thr185, p Tyr187]
• Immunogen: Phosphopeptide containing ERK1 T202/Y204 site
• Modification: p Tyr187, p Tyr204, p Thr185, p Thr202
• Specificity: Detects human, mouse, and rat ERK1 and ERK2 dually phosphorylated at T202/Y204 or T185/Y187, respectively.
• Source: N/A
• Isotype: IgG
• Clonality: Monoclonal
• Host: Rabbit

Applications/Dilutions

• Dilutions:
  • Western Blot 0.5 ug/mL
  • Simple Western 5 ug/mL
  • CyTOF-ready
  • Immunocytochemistry 8-25 ug/mL
  • Intracellular Staining by Flow Cytometry 2.5 ug/10^6 cells
• 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
• Reconstitution Instructions: Reconstitute at 0.5 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 Tyr187, p Tyr204, p Thr185, p Thr202] Antibody (269434) [Unconjugated]

• ERK1/ERK2

Background

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.

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 (MAB1018)(6)

Publications using MAB1018

• Rachel Barrow-McG Beta 1-integrin-c-Met cooperation reveals an inside-in survival signalling on autophagy-related endomembranes Nat Commun, 2016;7(0):11942. 2016 [PMID: 27336951]
• Clayton K, Haaland M, Douglas-Vail M, Mujib S, Chew G, Ndhlovu L, Ostrowski M T cell Ig and mucin domain-containing protein 3 is recruited to the immune synapse, disrupts stable synapse formation, and associates with receptor phosphatases. J Immunol, 2014;192(2):782-91. 2014 [PMID: 24337741] (ICC, Mouse)
• Shimizu T, Kasamatsu A, Yamamoto A, Koike K, Ishige S, Takatori H, Sakamoto Y, Ogawara K, Shiiba M, Tanzawa H, Uzawa K Annexin A10 in human oral cancer: biomarker for tumoral growth via G1/S transition by targeting MAPK signaling pathways. PLoS ONE, 2012;7(9):e45510. 2012 [PMID: 23029062] (Western Blot, Human)
• Lane AE, Tan JT, Hawkins CL The myeloperoxidase-derived oxidant HOSCN inhibits protein tyrosine phosphatases and modulates cell signalling via the mitogen-activated protein kinase (MAPK) pathway in macrophages. Biochem. J., 2010;430(1):161-9. 2010 [PMID: 20528774] (Western Blot, Mouse)
• Han Y, Wu G, Deng J, Tao J, Guo L, Tian X, Kang J, Zhang X, Yan C Cellular repressor of E1A-stimulated genes inhibits human vascular smooth muscle cell apoptosis via blocking P38/JNK MAP kinase activation. J. Mol. Cell. Cardiol., 2010;48(6):1225-35. 2010 [PMID: 20060003] (Western Blot, Human)
• Nevo I, Sagi-Assif O, Meshel T, Ben-Baruch A, Johrer K, Greil R, Trejo LE, Kharenko O, Feinmesser M, Yron I, Witz IP The involvement of the fractalkine receptor in the transmigration of neuroblastoma cells through bone-marrow endothelial cells. Cancer Lett., 2009;273(1):127-39. 2009 [PMID: 18778890] (Western Blot, Human)