Anti-Met (c-Met) antibody [EP1454Y] (ab51067)

Overview

  • Product nameAnti-Met (c-Met) antibody [EP1454Y]
    See all Met (c-Met) primary antibodies
  • Description
    Rabbit monoclonal [EP1454Y] to Met (c-Met)
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Met (c-Met) aa 1-100 (N terminal).
    (Peptide available as ab167073)

  • Positive control
    • 293 cell lysate or human breast carcinoma tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Anti-Met (c-Met) antibody (HRP) [EP1454Y] (ab193599)

Properties

Applications

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
WB 1/1000 - 1/10000. Detects a band of approximately 160 kDa (predicted molecular weight: 156 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/100 - 1/250.
Flow Cyt 1/100 - 1/1000.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • FunctionReceptor for hepatocyte growth factor and scatter factor. Has a tyrosine-protein kinase activity. Functions in cell proliferation, scattering, morphogenesis and survival.
  • Involvement in diseaseNote=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 similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family.
    Contains 3 IPT/TIG domains.
    Contains 1 protein kinase domain.
    Contains 1 Sema domain.
  • DomainThe kinase domain is involved in SPSB1 binding.
  • Post-translational
    modifications
    Dephosphorylated by PTPRJ at Tyr-1349 and Tyr-1365.
  • Cellular localizationMembrane.
  • Information by UniProt
  • Database links
  • 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 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

Anti-Met (c-Met) antibody [EP1454Y] images



  • Predicted band size : 156 kDa

    Lane 1: Wild-type HAP1 cell lysate (40 µg)
    Lane 2: Met (c-Met) knockout HAP1 cell lysate (40 µg)
    Lane 3: HepG2 cell lysate (40µg) (40 µg)
    Lane 4: HEK293 cel lysate (40µg) (40 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab51067 observed at 240 kDa. Red - loading control, ab18058, observed at 124 kDa.

    ab51067 was shown to recognize Met (c-Met) when Met (c-Met) knockout samples were used, along with additional cross-reactive bands. Wild-type and Met (c-Met) knockout samples were subjected to SDS-PAGE. Ab51067 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • 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 counter-stained 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, and is shown in the inset.
  • All lanes : Anti-Met (c-Met) antibody [EP1454Y] (ab51067) at 1/2000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : HEK293 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Predicted band size : 156 kDa
    Observed band size : 190 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Overlay histogram showing HeLa cells fixed in 4% PFA and stained with purified ab51067 at a dilution of 1 in 100 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
  • Immunofluorescence staining of Jurkat cells with purified ab51067 at a working dilution of 1/100, counter-stained 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.

  • 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 counter-stained 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, and is shown in the inset.
  • Overlay histogram showing Jurkat cells stained with unpurified ab51067 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab51067, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluor® 488 (IgG, H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • Anti-Met (c-Met) antibody [EP1454Y] (ab51067) at 1/2000 dilution (unpurified) + 293 cell lysate at 10 µg

    Secondary
    Goat anti-Rabbit HRP labeled at 1/2000 dilution

    Predicted band size : 156 kDa
    Observed band size : 160 kDa (why is the actual band size different from the predicted?)
  • Unpurified ab51067 showing positive staining in Hepatocellular carcinoma tissue.

  • Unpurified ab51067 showing positive staining in Thyroid gland carcinoma tissue.

  • Unpurified ab51067 showing positive staining in Normal tonsil tissue.

References for Anti-Met (c-Met) antibody [EP1454Y] (ab51067)

This product has been referenced in:
  • Jia Y  et al. c-MET inhibition enhances the response of the colorectal cancer cells to irradiation in vitro and in vivo. Oncol Lett 11:2879-2885 (2016). WB ; Human . Read more (PubMed: 27073569) »
  • Matsumoto Y  et al. A phase II study of erlotinib monotherapy in pre-treated non-small cell lung cancer without EGFR gene mutation who have never/light smoking history: re-evaluation of EGFR gene status (NEJ006/TCOG0903). Lung Cancer 86:195-200 (2014). Human . Read more (PubMed: 25249428) »

See all 13 Publications for this product

Product Wall

Thank you for contacting us.
The isotype controls are as follows:
ab8158 -- ab18450 rat IgG2a
ab51067 -- ab125938 rabbit monoclonal
ab28364 -- ab27472 rabbit polyclonal
I hope this information is helpful to you. Please do not h...

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The anti-Met antibody ab51067 was tested on mouse tissue lysates (brain, heart, kidney and spleen) in western blotting, but the antibody has not been tested on mouse tissues by IHC.

Although it was negative with the above mouse tissue, ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH6
Sample Human Tissue sections (human gastric cancer xenograft)
Specification human gastric cancer xenograft
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative Formaldehyde
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Abcam user community

Verified customer

Submitted May 23 2013


The blocking peptide for ab51067 has been added to our catalog as ab167073.
http://www.abcam.com/index.html?datasheet=167073

I can confirm that ab51067 is sold as tissue culture supernatant. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody concentration is usual...

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As shown on the product datasheet http://www.abcam.com/ab51067 thisantibody is sold as tissue culture supernatant. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated ...

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Thank you for contacting Abcam.
The concentration for ab51067, lot #GR57435-3 is *********mg/mL.
Please let me know if there is anything else we can help you with.

Thank you for contacting us. The immunogen shares 94% identity with mouse cMet. However, we did not detect any bands in mouse tissue lysates (brain, heart, kidney, and spleen). Thus, I would not expect cross-reactivity with mouse. I hope this is helpfu...

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We guarantee all of our antibodies for 6 months if stored according to the datasheet. For the concentration, the exact concentration has not been determined but based on QC testing; it is believed that it is between 0.5-2mg/ml.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"