Human/Mouse Phospho-HGF R/c-MET (Y1234/Y1235) Antibody

Catalog # Availability Size / Price Qty
AF2480
AF2480-SP
Detection of HGF R/c‑MET in pervanadate-treated MCF‑7 Human Cell Line by Flow Cytometry.
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Citations (6)
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Human/Mouse Phospho-HGF R/c-MET (Y1234/Y1235) Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects human and mouse HGF R/c-MET when phosphorylated at Y1234/Y1235 in Western blots.
Source
Polyclonal Rabbit IgG
Purification
Antigen Affinity-purified
Immunogen
Phosphopeptide containing human HGF R/c-MET Y1234/1235 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
0.5 µg/mL
See below
Simple Western
5 µg/mL
See below
Immunohistochemistry
5-15 µg/mL
See below
CyTOF-reported
Brodie, T.M. et al. (2018) Cytometry PartA. 93: 406. Ready to be labeled using establishedconjugation methods. No BSA or other carrier proteins that could interfere withconjugation.
 
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
See below

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

Intracellular Staining by Flow Cytometry Detection of HGF R/c‑MET in pervanadate-treated MCF‑7 Human Cell Line by Flow Cytometry. View Larger

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.

Immunohistochemistry HGF R/c-MET in Mouse Embryo. View Larger

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.

Western Blot Detection of Human Phospho-HGF R/c-MET (Y1234/Y1235) by Western Blot. View Larger

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.

Simple Western Detection of Human Phospho-HGF R/c‑MET (Y1234/Y1235) by Simple WesternTM. View Larger

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

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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: HGF R/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.

References
  1. Birchmeier, C. et al. (2003) Nat. Rev. Mol. Cell Biol. 4:915.
  2. Corso, S. et al. (2005) Trends Mol. Med. 11:284.
  3. Gherardi, E. et al. (2003) Proc. Natl. Acad. Sci. 100:12039.
  4. Park, M. et al. (1987) Proc. Natl. Acad. Sci. 84:6379.
  5. Crepaldi, T. et al. (1994) J. Biol. Chem. 269:1750.
  6. Prat, M. et al. (1991) Mol. Cell. Biol. 12:5954.
  7. Rodrigues, G.A. et al. (1991) Mol. Cell. Biol. 11:2962.
  8. Kong-Beltran, M. et al. (2004) Cancer Cell 6:75.
  9. Naldini, L. et al. (1991) Mol. Cell. Biol. 11:1793.
  10. Ponzetto, C. et al. (1994) Cell 77:261.
  11. Jeffers, M. et al. (1997) Mol. Cell. Biol. 17:799.
  12. Orian-Rousseau, V. et al. (2002) Genes Dev. 16:3074.
  13. Klosek, S.K. et al. (2005) Biochem. Biophys. Res. Commun. 336:408.
  14. Jo, M. et al. (2000) J. Biol. Chem. 275:8806.
  15. Wang, X. et al. (2002) Mol. Cell 9:411.
  16. Trusolino, L. et al. (2001) Cell 107:643.
  17. Giordano, S. et al. (2002) Nat. Cell Biol. 4:720.
  18. Conrotto, P. et al. (2004) Oncogene 23:5131.
  19. Follenzi, A. et al. (2000) Oncogene 19:3041.
  20. Sonnenberg, E. et al. (1993) J. Cell Biol. 123:223.
Long Name
Hepatocyte Growth Factor Receptor
Entrez Gene IDs
4233 (Human); 17295 (Mouse)
Alternate Names
AUTS9; cMET; c-MET; EC 2.7.10; EC 2.7.10.1; hepatocyte growth factor receptor; HGF R; HGF receptor; HGF/SF receptor; HGFR; Met (c-Met); met proto-oncogene (hepatocyte growth factor receptor); met proto-oncogene tyrosine kinase; MET; oncogene MET; Proto-oncogene c-Met; RCCP2; Scatter factor receptor; SF receptor; Tyrosine-protein kinase Met

Product Datasheets

Citations for Human/Mouse Phospho-HGF R/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.

6 Citations: Showing 1 - 6
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  1. 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: WB
  2. 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 Paraffin-embedded
  3. 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
  4. 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: Protein Array Development
  5. 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 Paraffin-embedded
  6. 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 Frozen

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