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RabMAb

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

製品の概要

  • 製品名
    Anti-Met (c-Met) antibody [EP1454Y]
    Met (c-Met) 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP1454Y] to Met (c-Met)
  • アプリケーション
    適用あり: WB, IHC-P, ICC/IF, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    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)

  • ポジティブ・コントロール
    • 293 cell lysate or human breast carcinoma tissue.
  • 特記事項

    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.

    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)

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

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.

ターゲット情報

  • 機能
    Receptor for hepatocyte growth factor and scatter factor. Has a tyrosine-protein kinase activity. Functions in cell proliferation, scattering, morphogenesis and survival.
  • 関連疾患
    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.
  • 配列類似性
    Belongs to the protein kinase superfamily. Tyr protein kinase family.
    Contains 3 IPT/TIG domains.
    Contains 1 protein kinase domain.
    Contains 1 Sema domain.
  • ドメイン
    The kinase domain is involved in SPSB1 binding.
  • 翻訳後修飾
    Dephosphorylated by PTPRJ at Tyr-1349 and Tyr-1365.
  • 細胞内局在
    Membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像



  • 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 specifically recognize Met (c-Met) in wild-type HAP1 cells along with additional cross reactive bands. No bands were observed when Met (c-Met) knockout samples were used. 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 1/10,000 dilution respectively and incubated overnight at 4C. 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 1/10,000 dilution for 1hr 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.
  • 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.

  • 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).
  • All lanes : Anti-Met (c-Met) antibody [EP1454Y] (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
    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.

  • 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.



  • 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.

    This western blot image is a comparison between ab51067 and a competitor's top cited rabbit polyclonal antibody.

  • 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.

    See Abreview

  • 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

参考文献

This product has been referenced in:
  • Cho SH  et al. The Antitumor Effect of C-terminus of Hsp70-Interacting Protein via Degradation of c-Met in Small Cell Lung Cancer. Korean J Thorac Cardiovasc Surg 50:153-162 (2017). Read more (PubMed: 28593150) »
  • Wang H  et al. Establishment of patient-derived gastric cancer xenografts: a useful tool for preclinical evaluation of targeted therapies involving alterations in HER-2, MET and FGFR2 signaling pathways. BMC Cancer 17:191 (2017). Read more (PubMed: 28292264) »

See all 20 Publications for this product

レビューと Q&A

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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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Antigen Retrieval Buffer (100X Citrate Buffer pH 6.0) (ab94674)
Permeabilization
Yes - 0.05% tween 20
Specification
Breast
Blocking step
Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
Fixative
Formaldehyde
Username

Mr. David Ivancic

Verified customer

投稿 May 23 2017


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, ...

Read More
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
Username

Abcam user community

Verified customer

投稿 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"

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