Recombinant Anti-Met (c-Met) antibody [EPR19067] KO Tested (ab216574)

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 & Species

Application	Species
ELISA	Recombinant fragment
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
WB	Human Recombinant fragment

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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: A549 and HeLa cells.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19067
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human MET (Met (c-Met)) knockout HeLa cell line (ab265961)
KO cell lysates
- Human MET (c-Met) knockout HeLa cell lysate (ab256991)
Related Products
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
ELISA	Recombinant fragment
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
WB	Human Recombinant fragment

Application	Abreviews	Notes
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.
Flow Cyt		1/600.
ELISA		Use at an assay dependent concentration.

Notes
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.
Flow Cyt 1/600.
ELISA Use at an assay dependent concentration.

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
modifications

Dephosphorylated by PTPRJ at Tyr-1349 and Tyr-1365.
Cellular localization

Membrane.
Target information above from: UniProt accession P08581 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 4233 Human
- Omim: 164860 Human
- SwissProt: P08581 Human
- Unigene: 132966 Human
Alternative names
- AUTS9 antibody
- c met antibody
- D249 antibody
- Hepatocyte growth factor receptor antibody
- HGF antibody
- HGF receptor antibody
- HGF/SF receptor antibody
- HGFR antibody
- MET antibody
- Met proto oncogene antibody
- Met proto oncogene tyrosine kinase antibody
- MET proto oncogene, receptor tyrosine kinase antibody
- Met proto-oncogene (hepatocyte growth factor receptor) antibody
- Met proto-oncogene antibody
- Met protooncogene antibody
- MET_HUMAN antibody
- Oncogene MET antibody
- Par4 antibody
- Proto-oncogene c-Met antibody
- RCCP2 antibody
- Scatter factor receptor antibody
- SF receptor antibody
- Tyrosine-protein kinase Met antibody
see all

Western blot - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

All lanes : Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : MET knockout HAP1 whole cell lysate

Lysates/proteins at 40 µg per lane.

Predicted band size: 155 kDa

Lanes 1 - 2: Merged signal (red and green). Green - ab216574 observed at 155 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab216574 was shown to specifically recognize MET in wild-type HAP1 cells along with additional cross reactive bands. No bands were observed when MET knockout samples were used. Wild-type and MET knockout samples were subjected to SDS-PAGE. ab216574 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1hr at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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.
Immunocytochemistry/ Immunofluorescence - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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.
Western blot - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

All lanes : Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/1000 dilution

Lane 1 : A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

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: 45-175 kDa
why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

This antibody detects Pro-c-Met (175 kDa), c-Met beta subunit (145 kDa) [PMID: 22418436], c-Met alpha subunit (45 kDa) [PMID: 8710887] and a cleavage c-Met fragment (85 kDa) [PMID: 11786517].
Western blot - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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 seconds

Blocking/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].
Western blot - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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 second

Blocking and Diluting buffer and concentration: 5% NFDM /TBST
Western blot - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

All lanes : Anti-Met (c-Met) antibody [EPR19067] (ab216574) at 1/1000 dilution

Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa whole cell lysate treated with PNGase F

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

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.

After partial deglycosylation, the band of c-Met beta subunit (145 kDa) is shifted to 125 kDa. The 85 kDa glycosylated band is reduced and a ~60 kDa band appears.
ELISA - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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.
Immunocytochemistry/ Immunofluorescence - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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.
Flow Cytometry - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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.
Flow Cytometry - Anti-Met (c-Met) antibody [EPR19067] (ab216574)

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.
Anti-Met (c-Met) antibody [EPR19067] (ab216574)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet

Publishing research using ab216574? Please let us know so that we can cite the reference in this datasheet.

ab216574 has been referenced in 2 publications.

Shen Y et al. Endostar regulates EMT, migration and invasion of lung cancer cells through the HGF-Met pathway. Mol Cell Probes 45:57-64 (2019). PubMed: 31096000
Shi W et al. MACC-1 antibody target therapy suppresses growth and migration of non-small cell lung cancer. Mol Med Rep 16:7329-7336 (2017). PubMed: 28944826

Key features and details

Overview

Product name

Description

Host species

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

KO cell lines

KO cell lysates

Related Products

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Certificate of Compliance

References (2)

Customer reviews and Q&As

Post-translational
modifications