Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody (ab5662)
- Datasheet
- Specific References (9)
- Protocols
Overview
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Product nameAnti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody
See all Met (c-Met) primary antibodies -
DescriptionRabbit polyclonal to Met (c-Met) (phospho Y1230 + Y1234 + Y1235)
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Host speciesRabbit
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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.
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Tested applicationsSuitable for: IHC-P, ICC/IF, WBmore details
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Species reactivityReacts with: Mouse, Human
Predicted to work with: Rat
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Immunogen
Synthetic phosphopeptide derived from the region of human c-Met that contains tyrosines 1230, 1234 and 1235. (Peptide available as ab41697.)
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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).
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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
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FormLiquid
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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.
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Storage bufferPreservative: 0.05% Sodium Azide
Constituents: 50% Glycerol, PBS, 1mg/ml BSA -
Concentration information loading... -
PurityImmunogen affinity purified
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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.
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Primary antibody notesBinding 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).
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunohistochemistry kits
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 |
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| 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
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FunctionReceptor for hepatocyte growth factor and scatter factor. Has a tyrosine-protein kinase activity. Functions in cell proliferation, scattering, morphogenesis and survival.
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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.
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Post-translational
modificationsDephosphorylated by PTPRJ at Tyr-1349 and Tyr-1365. -
Cellular localizationMembrane.
- Information by UniProt
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Database links
- 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
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Alternative names
- AUTS9 antibody
- c met antibody
- D249 antibody
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Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody (ab5662)This image is courtesy of an abreview submitted by Dr Charles Pallangyo, MUHAS.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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody (ab5662)This image is courtesy of an abreview submitted by Thorsten Cramer, RWTH Aachen University, Germany.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.
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Western blot - Anti-Met (c-Met) (phospho Y1230 + Y1234 + Y1235) antibody (ab5662)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.
Protocols
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) »
- Umezu T et al. Replenishing exosomes from older bone marrow stromal cells with miR-340 inhibits myeloma-related angiogenesis. Blood Adv 1:812-823 (2017). Read more (PubMed: 29296725) »
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