Human/Mouse Phospho-HGFR/c-MET (Y1349) 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.
Scientific Data
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Detection of Human Phospho-HGF R/ c-MET (Y1349) by Western Blot. Western blot shows lysates of A431 human epithelial carcinoma 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 Phospho-HGF R/c-MET (Y1349) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3950), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-HGF R/c-MET (Y1349) 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 (Y1349) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3950) 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 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 (Y1349) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3950) 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.
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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- Klosek, S.K. et al. (2005) Biochem. Biophys. Res. Commun. 336:408.
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- Conrotto, P. et al. (2004) Oncogene 23:5131.
- Follenzi, A. et al. (2000) Oncogene 19:3041.
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Product Datasheets
Citations for Human/Mouse Phospho-HGFR/c-MET (Y1349) 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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HGF-independent regulation of MET and GAB1 by nonreceptor tyrosine kinase FER potentiates metastasis in ovarian cancer
Authors: Gaofeng Fan, Siwei Zhang, Yan Gao, Peter A. Greer, Nicholas K. Tonks
Genes & Development
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MET Receptor Tyrosine Kinase Inhibition Reduces Interferon-Gamma (IFN-gamma )-Stimulated PD-L1 Expression through the STAT3 Pathway in Melanoma Cells
Authors: Song, KY;Han, YH;Roehrich, H;Brown, ME;Torres-Cabala, C;Giubellino, A;
Cancers
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot
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