HGFR/c-MET Antibody [Unconjugated]

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 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 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 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.

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.

• Reactivity: Hu
• Applications: WB, Simple Western, Flow, IHC, B/N, CyTOF-ready, ICC/IF, KO
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

HGFR/c-MET Antibody [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 direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 30% cross-reactivity with recombinant mouse HGF R is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: MET
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Blockade of Receptor-ligand Interaction
  • CyTOF-ready
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Knockout Validated
  • Simple Western 25 ug/mL
  • Western Blot 0.1 ug/mL

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
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 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 [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, 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.

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 (AF276)(26)

Publications using AF276

• K Koo, A Wuenschman, A Rendahl, KY Song, C Forster, A Wolf-Ringw, A Borgatti, A Giubellino Expression and Prognostic Evaluation of the Receptor Tyrosine Kinase MET in Canine Malignant Melanoma Veterinary sciences, 2023-03-26;10(4):. 2023-03-26 [PMID: 37104404] (Western Blot, Canine)
• Zuo Q, Yang Y, Lyu Y, Yang C, Chen C, Salman S, Huang T, Wicks E, Jackson W, Datan E, Qin W, Semenza G Plexin-B3 expression stimulates MET signaling, breast cancer stem cell specification, and lung metastasis. Cell Rep, 2023-02-28;42(3):112164. 2023-02-28 [PMID: 36857181] (Western Blot, Human)
• BS Mesquita, MHAM Fens, A Di Maggio, EDC Bosman, WE Hennink, M Heger, S Oliveira The Impact of Nanobody Density on the Targeting Efficiency of PEGylated Liposomes International Journal of Molecular Sciences, 2022-11-29;23(23):. 2022-11-29 [PMID: 36499301] (Flow Cytometry, Human)
• Q Zhang, DK Jeppesen, JN Higginboth, R Graves-Dea, VQ Trinh, MA Ramirez, Y Sohn, AC Neininger, N Taneja, ET McKinley, H Niitsu, Z Cao, R Evans, SE Glass, KC Ray, WH Fissell, S Hill, KL Rose, WJ Huh, MK Washington, GD Ayers, DT Burnette, S Sharma, LH Rome, JL Franklin, YA Lee, Q Liu, RJ Coffey Supermeres are functional extracellular nanoparticles replete with disease biomarkers and therapeutic targets Nature Cell Biology, 2021-12-09;23(12):1240-1254. 2021-12-09 [PMID: 34887515] (Western Blot, Human)
• P Banerjee, GY Xiao, X Tan, VJ Zheng, L Shi, MNB Rabassedas, HF Guo, X Liu, J Yu, L Diao, J Wang, WK Russell, J Roszik, CJ Creighton, JM Kurie The EMT activator ZEB1 accelerates endosomal trafficking to establish a polarity axis in lung adenocarcinoma cells Nature Communications, 2021-11-03;12(1):6354. 2021-11-03 [PMID: 34732702] (ICC, Human)
• M Nászai, K Bellec, Y Yu, A Román-Fern, E Sandilands, J Johansson, AD Campbell, JC Norman, OJ Sansom, DM Bryant, JB Cordero RAL GTPases mediate EGFR-driven intestinal stem cell proliferation and tumourigenesis Elife, 2021-06-07;10(0):. 2021-06-07 [PMID: 34096503] (ELISA Capture, Human)
• C Fedeli, H Moreno, S Kunz The Role of Receptor Tyrosine Kinases in Lassa Virus Cell Entry Viruses, 2020-08-06;12(8):. 2020-08-06 [PMID: 32781509] (Flow Cytometry, Human)
• GY Xiao, SL Schmid FCHSD2 controls oncogenic ERK1/2 signaling outcome by regulating endocytic trafficking PLoS Biol., 2020-07-17;18(7):e3000778. 2020-07-17 [PMID: 32678845] (Flow Cytometry, Human)
• R Graves-Dea, G Bogatcheva, S Rehman, Y Lu, JN Higginboth, B Singh Broad-spectrum receptor tyrosine kinase inhibitors overcome de novo and acquired modes of resistance to EGFR-targeted therapies in colorectal cancer Oncotarget, 2019-02-12;10(13):1320-1333. 2019-02-12 [PMID: 30863492] (Western Blot, Human)
• D Novo, N Heath, L Mitchell, G Caligiuri, A MacFarlane, D Reijmer, L Charlton, J Knight, M Calka, E McGhee, E Dornier, D Sumpton, S Mason, A Echard, K Klinkert, J Secklehner, F Kruiswijk, K Vousden, IR Macpherson, K Blyth, P Bailey, H Yin, LM Carlin, J Morton, S Zanivan, JC Norman Mutant p53s generate pro-invasive niches by influencing exosome podocalyxin levels Nat Commun, 2018-11-29;9(1):5069. 2018-11-29 [PMID: 30498210] (Bioassay, Human)
• JB Xiao, XL Li, L Liu, G Wang, SN Hao, HJ Dong, XM Wang, YF Zhang, HD Liu The association of semaphorin 5A with lymph node metastasis and adverse prognosis in cervical cancer Cancer Cell Int., 2018-06-22;18(0):87. 2018-06-22 [PMID: 29977159] (Neutralization, Human)
• ML Scott, DT Coleman, KC Kelly, JL Carroll, B Woodby, WK Songock, JA Cardelli, JM Bodily Human papillomavirus type 16 E5-mediated upregulation of Met in human keratinocytes Virology, 2018-03-31;519(0):1-11. 2018-03-31 [PMID: 29609071] (ICC, Human)
• Rachel Barrow-McG Beta 1-integrin-c-Met cooperation reveals an inside-in survival signalling on autophagy-related endomembranes Nat Commun, 2016-06-23;7(0):11942. 2016-06-23 [PMID: 27336951] (Western Blot, Human)
• Deborah R Kaye Tumor and Plasma Met Levels in Non-Metastatic Prostate Cancer PLoS ONE, 2016-06-14;11(6):e0157130. 2016-06-14 [PMID: 27300295] (ELISA Development (Detection), Human)
• Zoratti G, Tanabe L, Varela F, Murray A, Bergum C, Colombo E, Lang J, Molinolo A, Leduc R, Marsault E, Boerner J, List K Targeting matriptase in breast cancer abrogates tumour progression via impairment of stromal-epithelial growth factor signalling. Nat Commun, 2015-04-15;6(0):6776. 2015-04-15 [PMID: 25873032] (IHC-P, Human)
• Tabernero J, Elez M, Herranz M, Rico I, Prudkin L, Andreu J, Mateos J, Carreras M, Han M, Gifford J, Credi M, Yin W, Agarwal S, Komarnitsky P, Baselga J A pharmacodynamic/pharmacokinetic study of ficlatuzumab in patients with advanced solid tumors and liver metastases. Clin Cancer Res, 2014-03-14;20(10):2793-804. 2014-03-14 [PMID: 24634378] (ELISA Development (Detection), Human)
• Hasenauer S, Malinger D, Koschut D, Pace G, Matzke A, von Au A, Orian-Rousseau V Internalization of Met requires the co-receptor CD44v6 and its link to ERM proteins. PLoS ONE, 2013-04-23;8(4):e62357. 2013-04-23 [PMID: 23626807] (ICC, Human)
• Muller , P A J, Trinidad , A G, Timpson , P, Morton , J P, Zanivan , S, van den Berghe , P V E, Nixon , C, Karim , S A, Caswell , P T, Noll , J E, Coffill , C R, Lane , D P, Sansom , O J, Neilsen , P M, Norman , J C, Vousden , K H Mutant p53 enhances MET trafficking and signalling to drive cell scattering and invasion. Oncogene, 2012-05-14;32(10):1252-65. 2012-05-14 [PMID: 22580601] (Western Blot, Human)
• Cho K, Park J, Park C, Lee D, Lee E, Kim D, Kim K, Yoon S, Park Y, Kim E, Cho S, Jang S, Park B, Chi S, Yoo S, Jang M, Kim H, Kim E, Jo K, Park Y Identification of a pivotal endocytosis motif in c-Met and selective modulation of HGF-dependent aggressiveness of cancer using the 16-mer endocytic peptide. Oncogene, 2012-04-23;32(8):1018-29. 2012-04-23 [PMID: 22525273] (Flow Cytometry, Human)
• Szabo R, Rasmussen AL, Moyer AB c-Met-induced epithelial carcinogenesis is initiated by the serine protease matriptase. Oncogene, 2011-01-10;30(17):2003-16. 2011-01-10 [PMID: 21217780] (IHC-P, Human)
• Abella JV, Parachoniak CA, Sangwan V, Park M Dorsal ruffle microdomains potentiate Met receptor tyrosine kinase signaling and down-regulation. J. Biol. Chem., 2010-06-07;285(32):24956-67. 2010-06-07 [PMID: 20529867] (ICC, Human)
• Xu L, Nilsson MB, Saintigny P, Cascone T, Herynk MH, Du Z, Nikolinakos PG, Yang Y, Prudkin L, Liu D, Lee JJ, Johnson FM, Wong KK, Girard L, Gazdar AF, Minna JD, Kurie JM, Wistuba II, Heymach JV Epidermal growth factor receptor regulates MET levels and invasiveness through hypoxia-inducible factor-1alpha in non-small cell lung cancer cells. Oncogene, 2010-02-15;29(18):2616-27. 2010-02-15 [PMID: 20154724] (IHC-P, Human)
• Goldoni S, Humphries A, Nystrom A, Sattar S, Owens RT, McQuillan DJ, Ireton K, Iozzo RV Decorin is a novel antagonistic ligand of the Met receptor. J. Cell Biol., 2009-05-11;185(4):743-54. 2009-05-11 [PMID: 19433454] (ICC, Human)
• Schmelzer E, Mutig K, Schrade P, Bachmann S, Gerlach JC, Zeilinger K Effect of human patient plasma ex vivo treatment on gene expression and progenitor cell activation of primary human liver cells in multi-compartment 3D perfusion bioreactors for extra-corporeal liver support. Biotechnol. Bioeng., 2009-07-01;103(4):817-27. 2009-07-01 [PMID: 19274748] (IHC-P, Human)
• Bogatkevich GS, Ludwicka-Bradley A, Highland KB, Hant F, Nietert PJ, Singleton CB, Feghali-Bostwick CA, Silver RM Impairment of the antifibrotic effect of hepatocyte growth factor in lung fibroblasts from African Americans: possible role in systemic sclerosis. Arthritis Rheum., 2007-07-01;56(7):2432-42. 2007-07-01 [PMID: 17599773] (Neutralization, Human)
• Skibinski G, Elborn JS, Ennis M Bronchial epithelial cell growth regulation in fibroblast cocultures: the role of hepatocyte growth factor. Am. J. Physiol. Lung Cell Mol. Physiol., 2007-03-23;293(1):L69-76. 2007-03-23 [PMID: 17384084] (Neutralization, Human)

Review for HGFR/c-MET Antibody (AF276) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Anonymous - WB Human 01/07/2015

Summary

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

Bioinformatics

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