Human Phospho-HGFR/c-MET (Y1003) Antibody

Catalog #: AF4059
2 Citations Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF4059
AF4059-SP

Detection of Human Phospho-HGF R/c-MET (Y1003) by Western Blot.

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All HGFR/c-MET Antibodies

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Citations (2)

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Human Phospho-HGFR/c-MET (Y1003) Antibody Summary

Species Reactivity

Human

Specificity

Detects human and mouse HGF R/c-MET when phosphorylated at Y1003.

Source

Polyclonal Rabbit IgG

Purification

Antigen Affinity-purified

Immunogen

Phosphopeptide containing human HGF R Y1003 site

Formulation

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.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

Immersion fixed paraffin-embedded sections of human breast cancer tissue

Immunocytochemistry

5-15 µg/mL

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.

Scientific Data

Western Blot

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Detection of Human Phospho-HGF R/c-MET (Y1003) 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 1 µg/mL of Rabbit Anti-Human Phospho-HGF R/c-MET (Y1003) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4059), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-HGF R/ c-MET (Y1003) at approximately 140 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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Phospho-HGF R/c-MET (Y1003) in A431 Human Cell Line. HGF R/c-MET phosphorylated at Y1003 was detected in immersion fixed A431 human epithelial carcinoma cell line untreated (lower panel) or treated (upper panel) with pervanadate using Rabbit Anti-Human Phospho-HGF R/c-MET (Y1003) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4059) 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). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

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: 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.

References

Birchmeier, C. et al. (2003) Nat. Rev. Mol. Cell Biol. 4:915.
Corso, S. et al. (2005) Trends Mol. Med. 11:284.
Gherardi, E. et al. (2003) Proc. Natl. Acad. Sci. 100:12039.
Park, M. et al. (1987) Proc. Natl. Acad. Sci. 84:6379.
Crepaldi, T. et al. (1994) J. Biol. Chem. 269:1750.
Prat, M. et al. (1991) Mol. Cell. Biol. 12:5954.
Rodrigues, G.A. et al. (1991) Mol. Cell. Biol. 11:2962.
Kong-Beltran, M. et al. (2004) Cancer Cell 6:75.
Naldini, L. et al. (1991) Mol. Cell. Biol. 11:1793.
Ponzetto, C. et al. (1994) Cell 77:261.
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.

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

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Product Datasheet

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Citations for Human Phospho-HGFR/c-MET (Y1003) 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.

2 Citations: Showing 1 - 2
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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., 2018-02-06;0(0):.
Species: Human
Sample Types: Cell Lysates
Applications: Protein Array
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., 2011-08-18;179(4):1657-66.
Species: Human
Sample Types: Tissue Homogenates
Applications: Array Development