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Signal Transduction Protein Phosphorylation Tyrosine Kinases Receptor Tyrosine Kinases
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Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) 抗体 (ab5662)

  • Datasheet
  • SDS
Reviews (7)Q&A (4)References (23)

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Western blot - Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody (ab5662)

    Key features and details

    • Rabbit polyclonal to Met (c-Met) (phospho Y1230 + Y1234 + Y1235)
    • Suitable for: WB
    • Reacts with: Human
    • Isotype: IgG

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    関連製品

    製品の概要

    • 製品名

      Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody
      Met (c-Met) 一次抗体 製品一覧
    • 製品の詳細

      Rabbit polyclonal to Met (c-Met) (phospho Y1230 + Y1234 + Y1235)
    • 由来種

      Rabbit
    • 特異性

      The phosphospecific antibody that has been generated does not distinguish between the dually (pYpY 1234/1235) and triply (pYpYpY1230/1234/1235) phosphorylated forms of c-Met, both of which are likely to represent activated forms of this receptor.
    • アプリケーション

      適用あり: WBmore details
    • 種交差性

      交差種: Human
      交差が予測される動物種: Mouse, Rat
    • 免疫原

      Synthetic phosphopeptide derived from the region of human c-Met that contains tyrosines 1230, 1234 and 1235. (Peptide available as ab41697.)

    • ポジティブ・コントロール

      • WB: HEK-293T whole cell lysate.
    • 特記事項

       

      Binding of scatter factor (SF)/hepatocyte growth factor (HGF) to the c Met receptor tyrosine kinase (RTK) triggers receptor dimerization and phosphorylation on multiple residues within the juxtamembrane, catalytic core and cytoplasmic tail domains, thereby regulating receptor internalization, catalytic activity and multisubstrate docking. c Met contains three tyrosines (Tyr-xx- x-Tyr-Tyr motif) within the activation loop of the catalytic domain. This is also seen with the insulin receptor, insulin-like growth factor receptor (IGF1) receptor and nerve growth factor (NGF) receptors/Trks, for which phosphorylation of all three tyrosines is required for full activation. With c Met (and the related family member, RON) phosphorylation of tyrosines 1234 and 1235 has been shown to be important in receptor activation. Activation of the c Met receptor results in binding and/or phosphorylation of many intracellular signaling proteins including multiple adaptor proteins (e.g., Grb2, Shc, Cbl, Crk, cortactin, paxillin, and GAB1), and a variety of other signal transducers (e.g., PI 3-kinase, FAK, Src, Erk1&2, JNK, PLC-ã, and STAT3).

      Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

      Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

      We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

      In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

      We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

      Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

      Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

    製品の特性

    • 製品の状態

      Liquid
    • 保存方法

      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.
    • バッファー

      pH: 7.30
      Preservative: 0.05% Sodium azide
      Constituents: PBS, 50% Glycerol, 0.1% BSA
    • Concentration information loading...
    • 精製度

      Immunogen affinity purified
    • 特記事項(精製)

      The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated c-Met protein. The final product is generated by affinity chromatography using a c Met-derived peptide that is phosphorylated at tyrosines 1230, 1234, 1235.
    • 一次抗体 備考

      Binding of scatter factor (SF)/hepatocyte growth factor (HGF) to the c Met receptor tyrosine kinase (RTK) triggers receptor dimerization and phosphorylation on multiple residues within the juxtamembrane, catalytic core and cytoplasmic tail domains, thereby regulating receptor internalization, catalytic activity and multisubstrate docking. c Met contains three tyrosines (Tyr-xx- x-Tyr-Tyr motif) within the activation loop of the catalytic domain. This is also seen with the insulin receptor, insulin-like growth factor receptor (IGF1) receptor and nerve growth factor (NGF) receptors/Trks, for which phosphorylation of all three tyrosines is required for full activation. With c Met (and the related family member, RON) phosphorylation of tyrosines 1234 and 1235 has been shown to be important in receptor activation. Activation of the c Met receptor results in binding and/or phosphorylation of many intracellular signaling proteins including multiple adaptor proteins (e.g., Grb2, Shc, Cbl, Crk, cortactin, paxillin, and GAB1), and a variety of other signal transducers (e.g., PI 3-kinase, FAK, Src, Erk1&2, JNK, PLC-ã, and STAT3).
    • ポリ/モノ

      ポリクローナル
    • アイソタイプ

      IgG
    • 研究分野

      • Signal Transduction
      • Protein Phosphorylation
      • Tyrosine Kinases
      • Receptor Tyrosine Kinases
      • Epigenetics and Nuclear Signaling
      • Transcription
      • Cancer susceptibility
      • Proto-oncogenes
      • Cancer
      • Oncoproteins/suppressors
      • Oncoproteins
      • Growth factor receptors

    関連製品

    • Compatible Secondaries

      • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
      • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Isotype control

      • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

    アプリケーション

    Our Abpromise guarantee covers the use of ab5662 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. Detects a band of approximately 169 kDa.

    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.
    • 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
    • 参照データベース

      • 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
      • Unigene: 86844 Mouse
      • Unigene: 10617 Rat
      see all
    • 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 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

    • Western blot - Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody (ab5662)
      Western blot - Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody (ab5662)
      All lanes : Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody (ab5662) at 1/1000 dilution

      Lane 1 : Unstimulated (-), HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell extract
      Lanes 2-5 : Stimulated (+) with HGF, HEK-293T whole cell extract

      Secondary
      All lanes : Goat F(ab’)2 antirabbit IgG HRP conjugate


      Peptide Competition:

       

      Prior primary antibody incubation:

      1 and 2 -  no peptide,

      3 - non-phosphopeptide corresponding to the immunogen,

      4 -generic phosphotyrosine-containing peptide),

      5 - phosphopeptide immunogen.

      SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C.

      Bands were detected using the Pierce SuperSignal method.

      The data show that only the phosphopeptide corresponding to c Met pYpYpY1230/1234/1235] block the antibody signal, demonstrating the specificity of the antibody.

      Note: There are three isoforms of c Met, two of which are recognized by this antibody.

    プロトコール

    • Western blot protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    データシートおよび資料

    • Datasheet
    • SDS
  • 参考文献 (23)

    ab5662 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab5662 は 23 報の論文で使用されています。

    • Ueki R  et al. A chemically unmodified agonistic DNA with growth factor functionality for in vivo therapeutic application. Sci Adv 6:eaay2801 (2020). PubMed: 32270033
    • Oka M  et al. Hepatocyte Growth Factor-Secreting Mesothelial Cell Sheets Suppress Progressive Fibrosis in a Rat Model of CKD. J Am Soc Nephrol 30:261-276 (2019). PubMed: 30635373
    • Imafuku A  et al. Rat Mesenchymal Stromal Cell Sheets Suppress Renal Fibrosis via Microvascular Protection. Stem Cells Transl Med 8:1330-1341 (2019). PubMed: 31675167
    • Bonan NF  et al. Inhibition of HGF/MET signaling decreases overall tumor burden and blocks malignant conversion in Tpl2-related skin cancer. Oncogenesis 8:1 (2019). PubMed: 30631034
    • Zhang J  et al. MicroRNA-206 exerts anti-oncogenic functions in esophageal squamous cell carcinoma by suppressing the c-Met/AKT/mTOR pathway. Mol Med Rep 19:1491-1500 (2019). PubMed: 30569129
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 11 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Mammary tissue)
    Permeabilization
    Yes - DAKO antigen retrieval
    Specification
    Mammary tissue
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 18 2018

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (Injured spinal cord)
    Antigen retrieval step
    None
    Permeabilization
    No
    Specification
    Injured spinal cord
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 26°C
    Fixative
    Paraformaldehyde
    Read More

    Dr. K. A.

    Verified customer

    投稿 Sep 19 2016

    Western blot abreview for Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (PC12 (Pheochromocytoma cell line))
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    10 µg
    Treatment
    Lane1 untreated Lane2 500μM CoCl2
    Specification
    PC12 (Pheochromocytoma cell line)
    Blocking step
    Block one p as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 24°C
    Read More

    Dr. K. A.

    Verified customer

    投稿 May 20 2015

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10 mM citrate 5 min in pressure cooker
    Sample
    Mouse Tissue sections (small intestine)
    Specification
    small intestine
    Permeabilization
    No
    Fixative
    10% Formalin
    Read More

    Mr. Thorsten Cramer

    Verified customer

    投稿 Sep 23 2014

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    BSA as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 3%
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate buffer
    Sample
    Mouse Tissue sections (colon)
    Specification
    colon
    Permeabilization
    No
    Fixative
    Paraformaldehyde
    Read More

    Dr. Charles Pallangyo

    Verified customer

    投稿 Jul 09 2013

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (lung)
    Specification
    lung
    Fixative
    Formaldehyde
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Read More

    Ms. Yu Fang Wang

    Verified customer

    投稿 Mar 18 2013

    Western blot abreview for Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody

    Inconclusive
    Abreviews
    Abreviews
    Application
    Western blot
    Sample
    Dog Cell lysate - whole cell (osteosarcoma)
    Loading amount
    30 µg
    Specification
    osteosarcoma
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jun 15 2012

    Question

    Thanks for your cooperation. I told our customer about your answers and our customer has some more questions. First, if the customer wants to use A431 cell instead of 293 T cell for positive control, does he need to transfect with human Met cDNA? Secondly, if he does not need to use transfected A431 cell, is Met in A431 cell phosphorylated by EGF? I think these questions are not easy to answer, too. But I hope I can get the answer as soon as possible because our customer already spent so long time for your last reply. Looking forward to your answer.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 13 2006

    Answer

    Thank you for your e-mail. I can clarify that both types of cells, 293T and A431 cells need to be transfected with human Met cDNA to express high detectable levels of the protein. Both types of cells need to be stimulated to show increased phosphorylation levels. I hope this clarification helps and have updated the datasheet too,

    Read More

    Abcam Scientific Support

    Answered on Jul 13 2006

    Question

    Our customer who bought this product has some qustion about your data for wastern blot. According to your western data on the datasheet, you got the band with 293T cell by treating HGF. Thus, our customer is wondering. First, how much protein or cell did you load for the western blot? Secondly, how long did you incubate 293 T cells in the serum free medium before treating HGF? The last, what was the dosage to get the band and how long did you treat it? The user would like to know about these things in detail. Please let ue know a proper answer as much as you can. Could you recommend me the good references about there questions? I look forward to your reply.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 28 2006

    Answer

    I was able to confirm the details of the stimulation for the ab5662 testing. An important detail about our testing is that we used 293T cells "transiently transfected with human Met cDNA prior to treatment". Please stress with the customer that these are "transfected cells". If the customer was attempting to replicate our testing with "untransfected" cells, they would not observe the same response. The cells were serum starved overnight prior to HGF treatment. The cells were treated with 100 ng/mL HGF for 15 minutes. We loaded 10 ug per mini-gel lane.

    Read More

    Abcam Scientific Support

    Answered on Jul 12 2006

    Question

    Do have the blocking peptide for sale?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 22 2005

    Answer

    Thank you for your email. Unfortunately the peptide is not available through Abcam. You may want to try our link to "The World's Antibody Gateway". The World's Antibody Gateway is a free search engine service provided by Abcam to help you to quickly find the antibodies that you are looking for. It is a free-text search engine developed by the Abcam team so that it searches the catalogues of all online antibody companies (currently 249).

    Read More

    Abcam Scientific Support

    Answered on Mar 22 2005

    1-10 of 11 Abreviews or Q&A

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