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

Overview

  • Product nameAnti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody
    See all Met (c-Met) primary antibodies
  • Description
    Rabbit polyclonal to Met (c-Met) (phospho Y1230 + Y1234 + Y1235)
  • SpecificityThe 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.
  • Tested applicationsIHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

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

  • Positive control
    • 293T human kidney cells transiently transfected with human Met cDNA, stimulated with HGF (100 ng/mL HGF for 15 minutes). Mouse myeloma (SP-1) cells stimulated with HGF. A431 cells transiently transfected with human Met cDNA, stimulated with HGF (100 ng/mL HGF for 15 minutes).
  • General notes

     

    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).

Properties

  • FormLiquid
  • Storage instructionsShipped 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 bufferPreservative: 0.05% Sodium Azide
    Constituents: 50% Glycerol, PBS, 1mg/ml BSA
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Purification notesThe 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.
  • Primary antibody notes 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).
  • Clonality Polyclonal
  • IsotypeIgG
  • Research Areas

Applications

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.

Application Abreviews Notes
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB 1/1000. Detects a band of approximately 169 kDa.

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
    • cmet antibody
    • D249 antibody
    • Hepatocyte growth factor receptor antibody
    • Hepatocyte growth factor receptor Precursor antibody
    • HGF antibody
    • HGF receptor antibody
    • HGF SF 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
    • Renal cell carcinoma papillary 2 gene antibody
    • Scatter factor receptor antibody
    • SF receptor antibody
    • Tyrosine-protein kinase Met antibody
    see all

Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody images

  • Peptide Competition: Extracts prepared from 293T cells left unstimulated (1) or stimulated with HGF (2-5) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with the ab5662 antibody, following prior incubation with: no peptide (1, 2), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphotyrosine-containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab’)2 antirabbit IgG HRP conjugate and 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.

References for Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody (ab5662)

This product has been referenced in:
  • Castoldi R  et al. Molecular characterization of novel trispecific ErbB-cMet-IGF1R antibodies and their antigen-binding properties. Protein Eng Des Sel 25:551-60 (2012). WB ; Human . Read more (PubMed: 22936109) »
  • Smotrov N  et al. Development of a cell-based assay for measurement of c-Met phosphorylation using AlphaScreen technology and high-content imaging analysis. J Biomol Screen 14:404-11 (2009). Read more (PubMed: 19403923) »

See all 2 Publications for this product

Product Wall

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
Username

Herr Prof. Thorsten Cramer

Verified customer

Submitted Sep 23 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Charles Pallangyo

Verified customer

Submitted Jul 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Ms. Yu Fang Wang

Verified customer

Submitted Mar 18 2013

Abcam has not validated the combination of species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Jun 15 2012

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 phosphoryla...

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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 thes...

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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 fi...

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Thank you for your enquiry. To our knowledge, this antibody has yet to be tested in this application. All tested applications are specified on Abcam product datasheets. If you decide to go ahead and purchase this product, please let us know how you get...

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