AF2480

antibody from R&D Systems

Targeting: MET DFNB97, HGFR, RCCP2

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

AF2480

Provider

R&D Systems

Product name

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

Antibody type

Polyclonal

Description

Antigen Affinity-purified. Detects human and mouse HGF R/c-MET when phosphorylated at Y1234/Y1235 in Western blots.

Reactivity

Human, Mouse

Host

Rabbit

Conjugate

Unconjugated

Isotype

IgG

Vial size

50 ug

Concentration

LYOPH

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.

References

Submitted references

Reviving oncogenic addiction to MET bypassed by BRAF (G469A) mutation.

Virzì AR, Gentile A, Benvenuti S, Comoglio PM
Proceedings of the National Academy of Sciences of the United States of America 2018 Oct 2;115(40):10058-10063

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Pulmonary pericytes regulate lung morphogenesis.

Kato K, Diéguez-Hurtado R, Park DY, Hong SP, Kato-Azuma S, Adams S, Stehling M, Trappmann B, Wrana JL, Koh GY, Adams RH
Nature communications 2018 Jun 22;9(1):2448

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Dual MET/EGFR therapy leads to complete response and resistance prevention in a MET-amplified gastroesophageal xenopatient cohort.

Apicella M, Migliore C, Capelôa T, Menegon S, Cargnelutti M, Degiuli M, Sapino A, Sottile A, Sarotto I, Casorzo L, Cassoni P, De Simone M, Comoglio PM, Marsoni S, Corso S, Giordano S
Oncogene 2017 Mar 2;36(9):1200-1210

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Targeting MET kinase with the small-molecule inhibitor amuvatinib induces cytotoxicity in primary myeloma cells and cell lines.

Phillip CJ, Zaman S, Shentu S, Balakrishnan K, Zhang J, Baladandayuthapani V, Taverna P, Redkar S, Wang M, Stellrecht CM, Gandhi V
Journal of hematology & oncology 2013 Dec 10;6:92

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Protein signatures for classification and prognosis of gastric cancer a signaling pathway-based approach.

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
The American journal of pathology 2011 Oct;179(4):1657-66

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Alternative proteolytic processing of hepatocyte growth factor during wound repair.

Buchstein N, Hoffmann D, Smola H, Lang S, Paulsson M, Niemann C, Krieg T, Eming SA
The American journal of pathology 2009 Jun;174(6):2116-28

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Combined inhibition of MET and EGFR suppresses proliferation of malignant mesothelioma cells.

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 Jul;30(7):1097-105

Western blot

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

Detection of Human Phospho-HGF R/c-MET (Y1234/Y1235) by Simple Western^TM. 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.

Immunocytochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

HGF R/c-MET in SH-4 Human Cell Line. HGF R/c-MET was detected in immersion fixed SH-4 human melanoma cell line stimulated with HGF (left panel; positive staining) and non-stimulated (right panel; negative staining) using Rabbit Anti-Human/Mouse Phospho-HGF R/c-MET (Y1234/Y1235) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2480) at 15 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). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

HGF R/c-MET in Human Renal Cell Carcinoma Tissue. HGF R/c-MET was detected in immersion fixed paraffin-embedded sections of human renal cell carcinoma tissue using Rabbit Anti-Human/Mouse Phospho-HGF R/c-MET (Y1234/Y1235) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2480) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). 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 cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Flow cytometry

Supportive validation

Submitted by

R&D Systems (provider)

Main image

Experimental details

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.