Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Canine

Applications

Validated:

Knockout Validated, Immunohistochemistry, Western Blot, Blockade of Receptor-ligand Interaction, Flow Cytometry, Immunocytochemistry, Simple Western, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Immunoprecipitation, Bioassay, ELISA Capture, ELISA Development (Detection)

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all HGFR/c-MET Primary Antibodies »

Product Specifications

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 direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 30% cross-reactivity with recombinant mouse HGF R is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human HGFR/c-MET Antibody

HGF R/c-MET antibody in HT-29 and U937 Human Cell Line by Immunocytochemistry (ICC).

HGF R/c‑MET in HT‑29 and U937 Human Cell Line.

HGF R/c-MET was detected in immersion fixed HT-29 human colon adenocarcinoma cell line (positive control, left panel) and U937 human histiocytic lymphoma cell line (negative control, right panel) using Goat Anti-Human HGF R/c-MET Antigen Affinity-purified Polyclonal Antibody (Catalog # AF276) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

HGF R/c-MET antibody in Human Liver by Immunohistochemistry (IHC-P).

HGF R/c‑MET in Human Liver.

HGF R/c-MET was detected in immersion fixed paraffin-embedded sections of human liver using Goat Anti-Human HGF R/c-MET Antigen Affinity-purified Polyclonal Antibody (Catalog # AF276) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

HGF R/c-MET antibody in Human Skin by Immunohistochemistry (IHC-P).

HGF R/c‑MET in Human Skin.

HGF R/c-MET was detected in immersion fixed paraffin-embedded sections of human skin using 15 µg/mL Goat Anti-Human HGF R/c-MET Antigen Affinity-purified Polyclonal Antibody (Catalog # AF276) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western Blot Shows Human HGF R/c-MET Antibody Specificity by Using Knockout Cell Line.

Western Blot Shows Human HGF R/c‑MET Specificity by Using Knockout Cell Line.

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HGF R/c-Met knockout HeLa cell line (KO). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human HGF R/c-MET Antigen Affinity-purified Polyclonal Antibody (Catalog # AF276) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). Specific bands were detected for HGF R/c-MET at approximately 150-200 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Human HGFR/c-MET by Simple WesternTM.

Simple Western lane view shows lysates of HepG2 human hepatocellular carcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for HGFR/c-MET at approximately 151 kDa (as indicated) using 25 µg/mL of Goat Anti-Human HGFR/c-MET Antigen Affinity-purified Polyclonal Antibody (Catalog # AF276). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of HGFR/c-MET in MDA-MB-231 cells by Flow Cytometry

MDA-MB-231 cells were stained with Goat Anti-Human HGFR/c-MET Antigen Affinity-purified Polyclonal Antibody (Catalog # AF276, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

Detection of HGFR/c-MET by Western Blot

Detection of HGFR/c-MET by Western Blot

The link between CD44v6&the cytoskeleton through Ezrin is required for Met internalization.(A) HeLa cells transfected with a rat CD44v4-7 construct deleted from the cytoplasmic domain (CD44v4-7 delta cyt) or a control vector. The kinetic of Met internalization upon HGF induction was measured in a MESNA experiment (Material&Methods). In the first sample cells kept at 4°C. –M refers to a sample obtained from cells that not treated with MESNA. The Western Blot analysis was performed with a Met specific antibody&the rat specific CD44v6 antibody 1.1ASML (Material&Methods). Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23626807), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human HGFR/c-MET by Western Blot

Detection of Human HGFR/c-MET by Western Blot

C. albicans induces c-Met, EGFR, and E-cadherin to form a multiprotein complex.(A-C) Proximity ligation assay showing the interaction of c-Met with E-cadherin, EGFR with E-cadherin, and c-Met with EGFR in oral epithelial cells with and without 20-min infection with C. albicans. Confocal microscopic images (A). Scale bar 10 μm. Signal counts (B). Proximity ligation assay showing the interaction of c-Met with E-cadherin, EGFR with E-cadherin, and c-Met with EGFR in oral epithelial cells with and without 20-min infection with C. albicans. (A) Confocal microscopic images. Scale bar 10 μm. (B) Signal counts. (C-E) Co-immunoprecipitation experiments in oral epithelial cells transfected with control or E-cadherin (E-cad) siRNA and then infected with C. albicans for 20 min. (C) Representative immunoblots of proteins obtained by immunoprecipitation with an anti-c-Met antibody. (D and E) Densitometric analysis of 5 immunoblots. Results are mean ± SD. **p < 0.01, ***p < 0.001, ****p < 0.0001, ns; not significant (two-tailed Student’s t test [B] or one-way ANOVA with Sidak’s multiple comparisons test [D and E]). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37611070), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human HGFR/c-MET by Western Blot

Detection of Human HGFR/c-MET by Western Blot

Functional interactions among c-Met, EGFR, and E-cadherin during the endocytosis of C. albicans.(A-C) Immunoblot analysis showing the effects of the EGFR inhibitor gefitinib and the c-Met inhibitor SGX523 on C. albicans-induced phosphorylation of c-Met and EGFR in oral epithelial cells after 20 min of infection. Representative immunoblot (A), densitometric analysis of 4 immunoblots showing the phosphorylation of EGFR (B) and c-Met (C). Data are mean ± SD. (D) Effects of SGX523 and gefitinib on the endocytosis of C. albicans by oral epithelial cells. (E and F) Endocytosis of C. albicans by NIH/3T3 cells that expressed human c-Met (E) or human c-Met, EGFR, and HER2 (F). (G) Confocal micrographs of NIH3T3 cells expressing no human receptors (control), human c-Met, human EGFR and HER2, or human c-Met, EGFR, and HER2. The cells were infected with wild-type C. albicans germ tubes for 20 min, after which phosphorylated c-Met was detected with a phosphospecific anti-c-Met antibody. Scale bar 10 μm. Results in (D-F) are the mean ± SD of three experiments, each performed in triplicate. *p < 05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns; not significant (one-way ANOVA with Sidak’s multiple comparisons test [B-D] or two-tailed Student’s t test [E and F]). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37611070), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of HGFR/c-MET by Western Blot

Detection of HGFR/c-MET by Western Blot

CD44v6 controls Met internalization.(A) Left side: HeLa cells were transfected either with a pool of control siRNAs or a mixture of two different CD44v6 specific siRNAs and starved for 24 hours (Material and Methods). The cells were biotinylated (0,5 mg/ml), induced with HGF (50 ng/ml) and treated with MESNA. After cell lysis, proteins were pulled down with a NeutrAvidin resin and subjected to Western Blot analysis with Met or the Transferrin receptor (TfR) antibodies. For the first sample the cells were kept at 4°C. –M: the cells were not treated with MESNA. Right side: Western Blot analysis of cell lysates of ctrl siRNAs and CD44v6 siRNAs transfected HeLa cells using the CD44v6 and the TfR antibodies. (B) Starved HeLa cells respectively HT29 cells were incubated with the v6 peptide or a control peptide for 10 minutes at 37°C and then induced with 25 ng/ml of HGF for the indicated time points. Cells were then either lysed and the lysates were subjected to Western Blot analysis for phospho-Met and Met (below) or cells were fixed, permeabilized and stained for Met with specific antibodies (red) (above). Nuclei were stained with Dapi and images were taken with a confocal microscope (Leica SPE) using a 63× objective. The quantification of three independent experiments (n = 40) is shown. The percentage of cells with Met exclusively located at the plasma membrane was calculated for each time point. Student´s t test: ***p<0,001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23626807), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human HGFR/c-MET by Western Blot

Detection of Human HGFR/c-MET by Western Blot

C. albicans activates c-Met in oral epithelial cells.(A) Confocal microscopic images of OKF6/TERT-2 oral epithelial cells infected with C. albicans SC5314 for 20 min. c-Met, the epidermal growth factor receptor (EGFR), and E-cadherin were detected by indirect immunofluorescence using specific antibodies. Arrows point to the organisms in the magnified insets. Scale bar 10 μm. (B and C) Immunoblot analysis showing the time course of the phosphorylation of c-Met and EGFR in oral epithelial cells induced by C. albicans germ tubes (B). Densitometric analysis of 4 immunoblots (C). Data are mean ± SD. (D and E) Knockdown of c-Met with siRNA (D) or inhibition of c-Met signaling with SG523 (E) in oral epithelial cells inhibits the endocytosis of C. albicans. (F) Stimulation of oral epithelial cells with recombinant hepatocyte growth factor (HGF) enhances the endocytosis of C. albicans. Data in (D-F) are the mean ± SD of three experiments, each performed in triplicate. **p < 0.01, ****p < 0.0001, ns; not significant (two-tailed Student’s t test [D and E] or one-way ANOVA with Sidak’s multiple comparisons test [F]). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37611070), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human HGFR/c-MET Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 0.5-2 µg/mL of this antibody will block 50% of the binding of 5 ng/mL of Recombinant Human HGF (Catalog # 294-HGN) to immobilized Recombinant Human HGF R/c-MET Fc Chimera (Catalog # 358‑MT) coated at 1 µg/mL (100 µL/well). At 10 μg/mL, this antibody will block >90% of the binding.

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: MDA‑MB‑231 human breast cancer cell line

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HT‑29 human colon adenocarcinoma cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human skin and human liver tissue

Knockout Validated

HGF R/c‑MET is specifically detected in HeLa human cervical epithelial carcinoma parental cell line but is not detectable in HGF R/c‑MET knockout HeLa cell line.

Simple Western

25 µg/mL
Sample: HepG2 human hepatocellular carcinoma cell line

Western Blot

0.1 µg/mL
Sample: Recombinant Human HGF R/c-MET Fc Chimera (Catalog # 358-MT)

Reviewed Applications

Read 1 review rated 4 using AF276 in the following applications:

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

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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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 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, 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. 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.

Long Name

Hepatocyte Growth Factor Receptor

Alternate Names

c-MET, cMET, HGF R, MET

Entrez Gene IDs

4233 (Human); 17295 (Mouse); 102123512 (Cynomolgus Monkey)

Gene Symbol

MET

UniProt

P08581

Additional HGFR/c-MET Products

Product Documents for Human HGFR/c-MET Antibody

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Product Specific Notices for Human HGFR/c-MET Antibody

For research use only

Citations for Human HGFR/c-MET Antibody

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 22897854
    Species: Human
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