Recombinant Anti-Met (c-Met) antibody [EPR19067] (ab216574)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19067] to Met (c-Met)
- Suitable for: WB, IHC-P, ICC/IF, Indirect ELISA, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Met (c-Met) antibody [EPR19067]
See all Met (c-Met) primary antibodies -
Description
Rabbit monoclonal [EPR19067] to Met (c-Met) -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, Indirect ELISA, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Human, Recombinant fragment -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: A549, HeLa and HepG2 whole cell lysates; Human liver lysate; 293T whole cell lysate transfected with a His-tagged human c-Met construct; HeLa whole cell lysate, untreated or treated with PNGase F. IHC-P: Human breast, colon, liver cancer and ovary cancer tissues. ICC/IF: HeLa and A549 cells. Flow Cyt (intra): A549 and HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19067 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab216574 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Detects a band of approximately 45-175 kDa (predicted molecular weight: 155 kDa).
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/1000.
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Indirect ELISA |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
1/600.
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Notes |
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WB
1/1000. Detects a band of approximately 45-175 kDa (predicted molecular weight: 155 kDa). |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/1000. |
Indirect ELISA
Use at an assay dependent concentration. |
Flow Cyt (Intra)
1/600. |
Target
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Function
Receptor for hepatocyte growth factor and scatter factor. Has a tyrosine-protein kinase activity. Functions in cell proliferation, scattering, morphogenesis and survival. -
Involvement in disease
Note=Activation of MET after rearrangement with the TPR gene produces an oncogenic protein.
Note=Defects in MET may be associated with gastric cancer.
Defects in MET are a cause of hepatocellular carcinoma (HCC) [MIM:114550].
Defects in MET are a cause of renal cell carcinoma papillary (RCCP) [MIM:605074]. It is a subtype of renal cell carcinoma tending to show a tubulo-papillary architecture formed by numerous, irregular, finger-like projections of connective tissue. Renal cell carcinoma is a heterogeneous group of sporadic or hereditary carcinoma derived from cells of the proximal renal tubular epithelium. It is subclassified into common renal cell carcinoma (clear cell, non-papillary carcinoma), papillary renal cell carcinoma, chromophobe renal cell carcinoma, collecting duct carcinoma with medullary carcinoma of the kidney, and unclassified renal cell carcinoma.
Note=A common allele in the promoter region of the MET shows genetic association with susceptibility to autism in some families. Functional assays indicate a decrease in MET promoter activity and altered binding of specific transcription factor complexes.
Note=MET activating mutations may be involved in the development of a highly malignant, metastatic syndrome known as cancer of unknown primary origin (CUP) or primary occult malignancy. Systemic neoplastic spread is generally a late event in cancer progression. However, in some instances, distant dissemination arises at a very early stage, so that metastases reach clinical relevance before primary lesions. Sometimes, the primary lesions cannot be identified in spite of the progresses in the diagnosis of malignancies. -
Sequence similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family.
Contains 3 IPT/TIG domains.
Contains 1 protein kinase domain.
Contains 1 Sema domain. -
Domain
The kinase domain is involved in SPSB1 binding. -
Post-translational
modificationsDephosphorylated by PTPRJ at Tyr-1349 and Tyr-1365. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 4233 Human
- Omim: 164860 Human
- SwissProt: P08581 Human
- Unigene: 132966 Human
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Alternative names
- AUTS9 antibody
- c met antibody
- D249 antibody
see all
Images
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All lanes : Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/1000 dilution
All lanes :
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 155 kDa
Observed band size: 156 kDa why is the actual band size different from the predicted?Lanes 1 - 2: Merged signal (red and green). Green - ab216574 observed at 156 kDa. Red - loading control, ab8245 observed at 37 kDa.
ab216574 was shown to react with Met (c-Met) in wild-type HeLa. Loss of signal was observed when knockout cell line ab265961 (knockout cell lysate ab256991) was used. Wild-type and Met (c-Met) knockout samples were subjected to SDS-PAGE. ab216574 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Met (c-Met) with ab216574 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on HeLa cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/1000 dilution + Human liver lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 155 kDa
Observed band size: 100-150 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
In human liver the antibody detected c-Met beta subunit (145 kDa) [PMID: 22418436] and a cleavage c-Met fragment (100 kDa) [PMID: 18187039].
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All lanes : Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/10000 dilution
Lane 1 : 293T whole cell lysate (Human epithelial cell line from embryonic kidney) transfected with an empty expression vector
Lane 2 : 293T whole cell lysate transfected with a His-tagged human c-Met construct
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 155 kDa
Observed band size: 150-175 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking and Diluting buffer and concentration: 5% NFDM /TBST
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Immunohistochemical analysis of paraffin-embedded human breast tissue labeling Met (c-Met) with ab216574 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Membranous staining on human breast is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ELISA analysis of Human c-met recombinant protein at 1000 ng/mL with ab216574. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
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Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Met (c-Met) with ab216574 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Membranous staining on human colon is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human liver cancer tissue labeling Met (c-Met) with ab216574 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Membranous staining on tumor cells of human liver cancer is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue labeling Met (c-Met) with ab216574 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Cytoplasmic and membranous staining on tumor cells of human ovary cancer is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A549 (Human lung carcinoma cell line) cells labeling Met (c-Met) with ab216574 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasmic staining on A549 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed A549 (Human lung carcinoma cell line) cells labeling Met (c-Met) with ab216574 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Met (c-Met) with ab216574 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (11)
ab216574 has been referenced in 11 publications.
- Jin Y et al. A Novel c-MET-Targeting Antibody-Drug Conjugate for Pancreatic Cancer. Front Oncol 11:634881 (2021). PubMed: 33816276
- Lüttich L et al. Tyrosine Kinase c-MET as Therapeutic Target for Radiosensitization of Head and Neck Squamous Cell Carcinomas. Cancers (Basel) 13:N/A (2021). PubMed: 33919702
- Gu YH et al. Combined BRM270 and endostatin inhibit relapse of NSCLC while suppressing lung cancer stem cell proliferation induced by endostatin. Mol Ther Oncolytics 22:565-573 (2021). PubMed: 34553041
- Karmacharya U et al. Novel Pyridine Bioisostere of Cabozantinib as a Potent c-Met Kinase Inhibitor: Synthesis and Anti-Tumor Activity against Hepatocellular Carcinoma. Int J Mol Sci 22:N/A (2021). PubMed: 34575841
- Hyun SW et al. The sialidase NEU1 directly interacts with the juxtamembranous segment of the cytoplasmic domain of mucin-1 to inhibit downstream PI3K-Akt signaling. J Biol Chem 297:101337 (2021). PubMed: 34688655