Human/Mouse Phospho-HGFR/c-MET (Y1234/Y1235) Antibody
Human/Mouse Phospho-HGFR/c-MET (Y1234/Y1235) Antibody Summary
Applications
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
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Detection of Human Phospho-HGF R/c-MET (Y1234/Y1235) by Western Blot. Western blot shows Goat Anti-Human HGF R/c-MET Antigen Affinity-purified Polyclonal Antibody (Catalog # AF276) immunoprecipitate of MDA‑MB‑468 human breast cancer cell line untreated (-) or treated (+) with 100 μM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse Phospho-HGF R/c‑MET (Y1234/Y1235) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2480), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-HGF R/c‑MET (Y1234/Y1235) at approximately 145 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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HGF R/c‑MET in SH-4 Human Cell Line. HGF R/c‑MET was detected in immersion fixed SH-4 human melanoma cell line stimulated with HGF (left panel; positive staining) and non-stimulated (right panel; negative staining) using Rabbit Anti-Human/Mouse Phospho-HGF R/c‑MET (Y1234/Y1235) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2480) at 15 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.
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HGF R/c-MET in Mouse Embryo. HGF R/c-MET was detected in immersion fixed frozen sections of mouse embryo (13 d.p.c.) using Rabbit Anti-Human/Mouse Phospho-HGF R/c-MET (Y1234/Y1235) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2480) at 15 µg/mL overnight at 4 °C. Tissue was stained using 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.
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HGF R/c‑MET in Human Renal Cell Carcinoma Tissue. HGF R/c‑MET was detected in immersion fixed paraffin-embedded sections of human renal cell carcinoma tissue using Rabbit Anti-Human/Mouse Phospho-HGF R/c‑MET (Y1234/Y1235) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2480) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
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Detection of HGF R/c‑MET in pervanadate-treated MCF‑7 Human Cell Line by Flow Cytometry. MCF‑7 human breast cancer cell line was unstimulated (light orange open histogram) or treated with 100 μM pervanadate for 10 minutes (dark orange filled histogram), then stained with Rabbit Anti-Human/Mouse Phospho-HGF R/c‑MET (Y1234/Y1235) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF2480), or control antibody (Catalog # AB‑105‑C, blue open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.
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Detection of Human Phospho-HGF R/c‑MET (Y1234/Y1235) by Simple WesternTM. Simple Western lane view shows lysates of MDA‑MB‑468 human breast cancer cell line untreated (-) or treated (+) with 100 µM Pervanadate (PV) for 10 minutes, loaded at 0.2 mg/mL. A specific band was detected for HGF R/c‑MET at approximately 156 kDa (as indicated) using 5 µg/mL of Rabbit Anti-Human/Mouse Phospho-HGF R/c‑MET (Y1234/Y1235) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2480). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Reconstitution Calculator
Preparation and Storage
- 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: HGFR/c-MET
HGF R, also known as Met (from N-methyl-N’-nitro-N-nitrosoguanidine induced), is a glycosylated receptor tyrosine kinase that plays a central role in epithelial morphogenesis and cancer development. HGF R is synthesized as a single chain precursor which undergoes cotranslational proteolytic cleavage. This generates a mature HGF R that is a disulfide-linked dimer composed of a 50 kDa extracellular alpha chain and a 145 kDa transmembrane beta chain (1, 2). The extracellular domain (ECD) contains a seven bladed beta -propeller sema domain, a cysteine-rich PSI/MRS, and four Ig-like E-set domains, while the cytoplasmic region includes the tyrosine kinase domain (3, 4). Proteolysis and alternate splicing generate additional forms of human HGF R which either lack of the kinase domain, consist of secreted extracellular domains, or are deficient in proteolytic separation of the alpha and beta chains (5‑7). The sema domain, which is formed by both the alpha and beta chains of HGF R, mediates both ligand binding and receptor dimerization (3, 8). Ligand-induced tyrosine phosphorylation in the cytoplasmic region activates the kinase domain and provides docking sites for multiple SH2-containing molecules (9, 10). HGF stimulation induces HGF R downregulation via internalization and proteasome-dependent degradation (11). In the absence of ligand, HGF R forms noncovalent complexes with a variety of membrane proteins including CD44v6, CD151, EGF R, Fas, Integrin alpha 6/ beta 4, Plexins B1, 2, 3, and MSP R/Ron (12‑19). Ligation of one complex component triggers activation of the other, followed by cooperative signaling effects (12‑19). Formation of some of these heteromeric complexes is a requirement for epithelial cell morphogenesis and tumor cell invasion (12, 16, 17). Paracrine induction of epithelial cell scattering and branching tubulogenesis results from the stimulation of HGF R on undifferentiated epithelium by HGF released from neighboring mesenchymal cells (20). Genetic polymorphisms, chromosomal translocation, overexpression, and additional splicing and proteolytic cleavage of HGF R have been described in a wide range of cancers (1). Within the ECD, human HGF R shares 86%‑88% aa sequence identity with canine, mouse, and rat HGF R.
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- Jeffers, M. et al. (1997) Mol. Cell. Biol. 17:799.
- Orian-Rousseau, V. et al. (2002) Genes Dev. 16:3074.
- Klosek, S.K. et al. (2005) Biochem. Biophys. Res. Commun. 336:408.
- Jo, M. et al. (2000) J. Biol. Chem. 275:8806.
- Wang, X. et al. (2002) Mol. Cell 9:411.
- Trusolino, L. et al. (2001) Cell 107:643.
- Giordano, S. et al. (2002) Nat. Cell Biol. 4:720.
- Conrotto, P. et al. (2004) Oncogene 23:5131.
- Follenzi, A. et al. (2000) Oncogene 19:3041.
- Sonnenberg, E. et al. (1993) J. Cell Biol. 123:223.
Product Datasheets
Citations for Human/Mouse Phospho-HGFR/c-MET (Y1234/Y1235) Antibody
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.
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Reviving oncogenic addiction to MET bypassed by BRAF (G469A) mutation
Authors: AR Virzì, A Gentile, S Benvenuti, PM Comoglio
Proc. Natl. Acad. Sci. U.S.A., 2018;0(0):.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC -
Pulmonary pericytes regulate lung morphogenesis
Authors: K Kato, R Diéguez-Hu, DY Park, SP Hong, S Kato-Azuma, S Adams, M Stehling, B Trappmann, JL Wrana, GY Koh, RH Adams
Nat Commun, 2018;9(1):2448.
Species: Mouse
Sample Types: Cell Lysates
Applications: Western Blot -
Antiproliferative effects of the CDK6 inhibitor PD0332991 and its effect on signaling networks in gastric cancer cells
Authors: D Wang, Y Sun, W Li, F Ye, Y Zhang, Y Guo, DY Zhang, J Suo
Int. J. Mol. Med., 2018;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Protein Array -
Dual MET/EGFR therapy leads to complete response and resistance prevention in a MET-amplified gastroesophageal xenopatient cohort
Oncogene, 2016;0(0):.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Targeting MET kinase with the small-molecule inhibitor amuvatinib induces cytotoxicity in primary myeloma cells and cell lines.
Authors: Phillip C, Zaman S, Shentu S, Balakrishnan K, Zhang J, Baladandayuthapani V, Taverna P, Redkar S, Wang M, Stellrecht C, Gandhi V
J Hematol Oncol, 2013;6(0):92.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Protein signatures for classification and prognosis of gastric cancer a signaling pathway-based approach.
Authors: Wang D, Ye F, Sun Y, Li W, Liu H, Jiang J, Zhang Y, Liu C, Tong W, Gao L, Sun Y, Zhang W, Seetoe T, Lee P, Suo J, Zhang DY
Am. J. Pathol., 2011;179(4):1657-66.
Species: Human
Sample Types: Tissue Homogenates
Applications: Array Development -
Combined inhibition of MET and EGFR suppresses proliferation of malignant mesothelioma cells.
Authors: Kawaguchi K, Murakami H, Taniguchi T, Fujii M, Kawata S, Fukui T, Kondo Y, Osada H, Usami N, Yokoi K, Ueda Y, Yatabe Y, Ito M, Horio Y, Hida T, Sekido Y
Carcinogenesis, 2009;30(7):1097-105.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-P -
Alternative proteolytic processing of hepatocyte growth factor during wound repair.
Authors: Buchstein N, Hoffmann D, Smola H, Lang S, Paulsson M, Niemann C, Krieg T, Eming SA
Am. J. Pathol., 2009;174(6):2116-28.
Species: Human
Sample Types: Whole Tissue
Applications: IHC-Fr
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