Recombinant Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1454Y] to Met (c-Met) - N-terminal
- Suitable for: ELISA, WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-Met (c-Met) antibody [EP1454Y] - N-terminal
See all Met (c-Met) primary antibodies -
Description
Rabbit monoclonal [EP1454Y] to Met (c-Met) - N-terminal -
Host species
Rabbit -
Tested applications
Suitable for: ELISA, WB, IHC-P, ICC/IFmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse, Rat, Human, Recombinant fragment -
Immunogen
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Positive control
- WB: Wild-type HAP1 cell lysate. HepG2, HEK-293 and HeLa cell lysate. Mouse and rat thymus tissue lysate. Mouse lung tissue lysate. Hela and A459 whole cell lysate. IHC-P: Human bladder carcinoma and clear cell kidney carcinoma tissue. ICC/IF: Jurkat 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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1454Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab51067 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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ELISA |
Use at an assay dependent concentration.
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WB |
1/1000 - 1/10000. Detects a band of approximately 160 kDa (predicted molecular weight: 156 kDa).Can be blocked with Human Met (c-Met) peptide (ab167073).
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IHC-P | (10) |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF | (1) |
1/100 - 1/250.
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Notes |
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ELISA
Use at an assay dependent concentration. |
WB
1/1000 - 1/10000. Detects a band of approximately 160 kDa (predicted molecular weight: 156 kDa).Can be blocked with Human Met (c-Met) peptide (ab167073). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/250. |
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
- Entrez Gene: 17295 Mouse
- Entrez Gene: 24553 Rat
- Omim: 164860 Human
- SwissProt: P08581 Human
- SwissProt: P16056 Mouse
- SwissProt: P97523 Rat
- Unigene: 132966 Human
see all -
Alternative names
- AUTS9 antibody
- c met antibody
- D249 antibody
see all
Images
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Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067) at 1/1000 dilution + HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Predicted band size: 156 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?
Exposure time: 60 secondsBlocking buffer / Diluent and concentration:
5% NFDM/TBST
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All lanes : Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : MET knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 156 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Met (c-Met) antibody [EP1454Y] - N-terminal staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab51067 was shown to bind specifically to the alpha chain of c-Met. A band was observed at 50 kDa in wild-type HeLa cell lysates with no signal observed at this size in MET knockout cell line ab265961 (knockout cell lysate ab256991). To generate this image, wild-type and MET knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
Immunohistochemical staining of paraffin embedded human bladder carcinoma with purified ab51067 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.
PBS was used instead of the primary antibody as the negative control (inset).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)This image is courtesy of an Abreview submitted by David Ivancic.
ab51067 staining Met (c-Met) in human breast tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).
Tissue was fixed with formaldehyde, permeabilized with 0.05% Tween-20 and blocked for 30 minutes at 22°C; antigen retrieval was by heat mediation in antigen retrieval buffer (100X citrate buffer pH 6.0) (ab94674). Samples were incubated with the primary antibody (1/100) for 14 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/300) was used as the secondary antibody.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
Immunohistochemical staining of paraffin embedded human clear cell kidney carcinoma with purified ab51067 at a working dilution of 1/100. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.
PBS was used instead of the primary antibody as the negative control (inset).
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All lanes : Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067) at 1/1000 dilution
Lane 1 : Mouse thymus tissue lysate
Lane 2 : Rat thymus tissue lysate
Lane 3 : Mouse lung tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 156 kDa
Observed band size: 240 kDa why is the actual band size different from the predicted?Lanes 1 - 3: Merged signal (red and green). Green - ab51067 observed at 240 kDa. Red - loading control, ab18058, observed at 130 kDa.
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab51067 and ab18058 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.
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ELISA analysis of Human Met recombinant protein at 1000 ng/mL with ab51067. 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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Immunocytochemistry/ Immunofluorescence - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (ab51067)
Immunofluorescence staining of Jurkat (Human T cell leukemia cell line from peripheral blood) cells with purified ab51067 at a working dilution of 1/100, counterstained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel.
The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100.
The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab51067 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (87)
ab51067 has been referenced in 87 publications.
- Wu X et al. Down-regulation of the tumor suppressor miR-34a contributes to head and neck cancer by up-regulating the MET oncogene and modulating tumor immune evasion. J Exp Clin Cancer Res 40:70 (2021). PubMed: 33596979
- Li M et al. Knockdown of SLC39A4 Expression Inhibits the Proliferation and Motility of Gallbladder Cancer Cells and Tumor Formation in Nude Mice. Cancer Manag Res 13:2235-2246 (2021). PubMed: 33727860
- Chen JX et al. TRIM47 promotes malignant progression of renal cell carcinoma by degrading P53 through ubiquitination. Cancer Cell Int 21:129 (2021). PubMed: 33622324
- Stewart CM et al. Ebola virus triggers receptor tyrosine kinase-dependent signaling to promote the delivery of viral particles to entry-conducive intracellular compartments. PLoS Pathog 17:e1009275 (2021). PubMed: 33513206
- Pereira PMR et al. Immuno-PET Detects Changes in Multi-RTK Tumor Cell Expression Levels in Response to Targeted Kinase Inhibition. J Nucl Med 62:366-371 (2021). PubMed: 32646879