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

Overview

  • Product name
    Anti-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)
  • Specificity
    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.
  • Tested applications
    Suitable for: IHC-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

  • 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
    Preservative: 0.05% Sodium Azide
    Constituents: 50% Glycerol, PBS, 1mg/ml BSA
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    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.
  • 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
  • Isotype
    IgG
  • 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

  • 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.
  • 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

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse colon tissue sections labeling Met (c-Met) (phospho Y1230 + Y1234 + Y1235) with ab5662 at 1/100 dilution. the tissue was fixed with paraformaldehyde and heat-mediated antigen retrieval was performed using a citrate buffer. The sample was blocked with 3% BSA for 30 hours, followed by staining with ab5662 at 1/100 in 3% BSA/PBS for 12 hours at 4°C. An Alexa Fluor® 488 conjugated donkey anti-rabbit IgG secondary antibody was used at 1/1000 dilution.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse small intestine tissue sections labeling Met (c-Met) (phospho Y1230 + Y1234 + Y1235) with ab5662 at 1/200 dilution. The tissue was fixed in 10% formalin and heat-mediated antigen retrieval was performed using 10 mM citrate buffer for 5 minutes in a pressure cooker. A biotinylated goat anti-rabbit secondary antibody was used at 1/250 dilution.

    See Abreview

  • 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

This product has been referenced in:
  • Wang H  et al. DCLK1 is correlated with MET and ERK5 expression, and associated with prognosis in malignant pleural mesothelioma. Int J Oncol 51:91-103 (2017). Read more (PubMed: 28560410) »
  • Pajari AM  et al. Ellagitannin-rich cloudberry inhibits hepatocyte growth factor induced cell migration and phosphatidylinositol 3-kinase/AKT activation in colon carcinoma cells and tumors in Min mice. Oncotarget 7:43907-43923 (2016). IHC . Read more (PubMed: 27270323) »

See all 7 Publications for this product

Customer reviews and Q&As

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
Username

清隆 新井

Verified customer

Submitted Sep 19 2016

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
Username

清隆 新井

Verified customer

Submitted May 20 2015

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

Mr. 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

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"

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