HGFR/c-MET Antibody (85503) [Unconjugated]

HGF R/c-MET was detected in immersion fixed HT-29 human colon adenocarcinoma cell line using Mouse Anti-Human HGF R/c-MET Monoclonal Antibody (Catalog # MAB3581) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane and secreted molecule. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

HGF R/c‑MET was detected in immersion fixed paraffin-embedded sections of human renal cell carcinoma tissue using Mouse Anti-Human HGF R/c‑MET Monoclonal Antibody (Catalog # MAB3581) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). 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 plasma membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

ABT-700 specifically binds cellular c-Met and antagonizes c-Met signaling in both HGF-dependent and -independent settings. a FACS analysis of ABT-700 binding to MCF7 transfectants. Stable human c-Met or vector control transfectants of human MCF7 breast cancer cells were incubated with increasing amounts of ABT-700 and bound ABT-700 was detected by FACS with secondary anti-human IgG conjugated with Alexa 488. b ELISA quantification of phospho-c-Met in A549 cells. A549 cells grown in a 96-well plate were pre-incubated for one hour with antibodies in a dose-range as shown, followed by stimulation with 1 nM HGF for 10 min. Total cell lysates were made and phospho-c-Met was detected by ELISA. c ELISA quantification of phospho-c-Met in SNU5 cells. SNU5 cells grown in a 96-well plate were incubated with antibodies in a dose-range as shown for 6 h. Total cell lysates were made and subjected to ELISA for phospho-c-Met. The value of cells in media alone was used as 100 % of control. d ELISA quantification of total c-Met in SNU5 cells. SNU5 cells grown in a 96-well plate were incubated with antibodies in a dose-range as shown for 6 h. Total cell lysates were made and c-Met level was determined by ELISA. The value of cells in media alone was used as 100 % of control. e Western blot analysis of U87MG cell lysates. U87MG cells grown in a 12-well plate were treated with antibodies as shown at 10 μg/mL for 10 min, 1 h or 6 h. Total cell lysates were analyzed for c-Met and other phosphorylated targets as shown. Western blot analysis of Hs746T cell lysates. Hs746T cells grown in a 12-well pate were treated with antibodies as shown at 10 μg/mL for 6 h. Total cell lysates were analyzed for c-Met and other phosphorylated targets as shown. f Western blot analysis of SNU620 cell lysates. SNU620 cells grown in a 12-well pate were treated with antibodies as shown at 10 μg/mL for 24 h. Total cell lysates were analyzed for c-Met and other phosphorylated targets as shown. g Inhibition of proliferation of SNU620 cells. SNU620 cells were plated in a 96-well plate and treated with antibodies in a dose range as shown for 3 days. Quantification of live cells at the end of incubation was done with Cell-titer Glo reagents. The data shown in all panels are from one of at least two independent experiments showing similar results as described in Methods Image collected and cropped by CiteAb from the following publication (//www.biomedcentral.com/1471-2407/16/105), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: IHC, ELISA(Cap), ICC/IF
• Clone: 85503
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

HGFR/c-MET Antibody (85503) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human HGF R/c‑MET
  Glu25-Thr932
  Accession # P08581
• Specificity: Detects human HGF R/c‑MET in ELISAs. In sandwich immunoassays, no cross-reactivity with recombinant human (rh) EGFR or rhIGF-1 R is observed.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: MET
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • ELISA Capture (Matched Antibody Pair) 2-8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.1-0.4 ug/mL
  • ELISA Standard (Matched Pair)
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 5-25 ug/mL
• Application Notes: ELISA Detection: Human HGF R/c-MET Biotinylated Antibody (Catalog number BAF358)
  Standard: Recombinant Human HGF R/c-MET Fc Chimera (Catalog number 358-MT)

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 either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• 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 HGFR/c-MET Antibody (85503) [Unconjugated]

• 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

Background

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 non-covalent 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, over-expression, 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% amino acid sequence identity with canine, mouse, and rat HGF R.

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. USA 100:12039.
4. Park, M. et al. (1987) Proc. Natl. Acad. Sci. USA 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.

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 HGFR/c-MET Antibody (MAB3581)(4)

Publications using MAB3581

• Allison M. Claas, Lyla Atta, Simon Gordonov, Aaron S. Meyer, Douglas A. Lauffenburger Systems Modeling Identifies Divergent Receptor Tyrosine Kinase Reprogramming to MAPK Pathway Inhibition Cellular and Molecular Bioengineering 12/1/2018 [PMID: 30524510]
• Wang J, Goetsch L, Tucker L, Zhang Q, Gonzalez A, Vaidya K, Oleksijew A, Boghaert E, Song M, Sokolova I, Pestova E, Anderson M, Pappano W, Ansell P, Bhathena A, Naumovski L, Corvaia N, Reilly E Anti-c-Met monoclonal antibody ABT-700 breaks oncogene addiction in tumors with MET amplification. BMC Cancer, 2016-02-16;16(1):105. 2016-02-16 [PMID: 26879245] (ELISA Development, Human)
• Raimond Heukers, Isil Altintas, Smiriti Raghoenath, Erica De Zan, Richard Pepermans, Rob C. Roovers et al. Targeting hepatocyte growth factor receptor (Met) positive tumor cells using internalizing nanobody-decorated albumin nanoparticles Biomaterials 1/1/2014 [PMID: 24139763]
• Raimond Heukers, Isil Altintas, Smiriti Raghoenath, Erica De Zan, Richard Pepermans, Rob C. Roovers et al. Targeting hepatocyte growth factor receptor (Met) positive tumor cells using internalizing nanobody-decorated albumin nanoparticles Biomaterials 2014-01-01 [PMID: 24139763] (Western Blot, Human)

Bioinformatics

• Gene Symbol: MET
• Uniprot:
  • P08581()