Overview

  • Product nameAnti-TIE2 antibody [Cl. 16]
    See all TIE2 primary antibodies
  • Description
    Mouse monoclonal [Cl. 16] to TIE2
  • Tested applicationsSuitable for: IHC-Fr, IHC-FoFr, WB, ELISA, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment corresponding to Human TIE2. Recombinant human soluble extracellular TIE2.
    Database link: Q02763

Properties

Applications

Our Abpromise guarantee covers the use of ab24859 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-Fr 1/25. PubMed: 19253934
IHC-FoFr Use at an assay dependent concentration.
WB Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 126 kDa.
ELISA Use a concentration of 1 - 2 µg/ml.
Flow Cyt Use 1-2µg for 106 cells.

We tested in-house with methanol-fixed cells, but this may not be necessary since the antibody recognies an extracellular epitope.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. PubMed: 20622121

Target

  • FunctionTyrosine-protein kinase that acts as cell-surface receptor for ANGPT1, ANGPT2 and ANGPT4 and regulates angiogenesis, endothelial cell survival, proliferation, migration, adhesion and cell spreading, reorganization of the actin cytoskeleton, but also maintenance of vascular quiescence. Has anti-inflammatory effects by preventing the leakage of proinflammatory plasma proteins and leukocytes from blood vessels. Required for normal angiogenesis and heart development during embryogenesis. Required for post-natal hematopoiesis. After birth, activates or inhibits angiogenesis, depending on the context. Inhibits angiogenesis and promotes vascular stability in quiescent vessels, where endothelial cells have tight contacts. In quiescent vessels, ANGPT1 oligomers recruit TEK to cell-cell contacts, forming complexes with TEK molecules from adjoining cells, and this leads to preferential activation of phosphatidylinositol 3-kinase and the AKT1 signaling cascades. In migrating endothelial cells that lack cell-cell adhesions, ANGT1 recruits TEK to contacts with the extracellular matrix, leading to the formation of focal adhesion complexes, activation of PTK2/FAK and of the downstream kinases MAPK1/ERK2 and MAPK3/ERK1, and ultimately to the stimulation of sprouting angiogenesis. ANGPT1 signaling triggers receptor dimerization and autophosphorylation at specific tyrosine residues that then serve as binding sites for scaffold proteins and effectors. Signaling is modulated by ANGPT2 that has lower affinity for TEK, can promote TEK autophosphorylation in the absence of ANGPT1, but inhibits ANGPT1-mediated signaling by competing for the same binding site. Signaling is also modulated by formation of heterodimers with TIE1, and by proteolytic processing that gives rise to a soluble TEK extracellular domain. The soluble extracellular domain modulates signaling by functioning as decoy receptor for angiopoietins. TEK phosphorylates DOK2, GRB7, GRB14, PIK3R1; SHC1 and TIE1.
  • Tissue specificityDetected in umbilical vein endothelial cells. Proteolytic processing gives rise to a soluble extracellular domain that is detected in blood plasma (at protein level). Predominantly expressed in endothelial cells and their progenitors, the angioblasts. Has been directly found in placenta and lung, with a lower level in umbilical vein endothelial cells, brain and kidney.
  • Involvement in diseaseDominantly inherited venous malformations
    May play a role in a range of diseases with a vascular component, including neovascularization of tumors, psoriasis and inflammation.
  • Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. Tie subfamily.
    Contains 3 EGF-like domains.
    Contains 3 fibronectin type-III domains.
    Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 protein kinase domain.
  • DomainThe soluble extracellular domain is functionally active in angiopoietin binding and can modulate the activity of the membrane-bound form by competing for angiopoietins.
  • Post-translational
    modifications
    Proteolytic processing leads to the shedding of the extracellular domain (soluble TIE-2 alias sTIE-2).
    Autophosphorylated on tyrosine residues in response to ligand binding. Autophosphorylation occurs in trans, i.e. one subunit of the dimeric receptor phosphorylates tyrosine residues on the other subunit. Autophosphorylation occurs in a sequential manner, where Tyr-992 in the kinase activation loop is phosphorylated first, followed by autophosphorylation at Tyr-1108 and at additional tyrosine residues. ANGPT1-induced phosphorylation is impaired during hypoxia, due to increased expression of ANGPT2. Phosphorylation is important for interaction with GRB14, PIK3R1 and PTPN11. Phosphorylation at Tyr-1102 is important for interaction with SHC1, GRB2 and GRB7. Phosphorylation at Tyr-1108 is important for interaction with DOK2 and for coupling to downstream signal transduction pathways in endothelial cells. Dephosphorylated by PTPRB.
    Ubiquitinated. The phosphorylated receptor is ubiquitinated and internalized, leading to its degradation.
  • Cellular localizationCell membrane. Cell junction. Cell junction, focal adhesion. Cytoplasm, cytoskeleton. Secreted. Recruited to cell-cell contacts in quiescent endothelial cells. Colocalizes with the actin cytoskeleton and at actin stress fibers during cell spreading. Recruited to the lower surface of migrating cells, especially the rear end of the cell. Proteolytic processing gives rise to a soluble extracellular domain that is secreted.
  • Information by UniProt
  • Database links
  • Alternative names
    • Angiopoietin 1 receptor antibody
    • Angiopoietin-1 receptor antibody
    • CD202b antibody
    • CD202b antigen antibody
    • Endothelial tyrosine kinase antibody
    • Endothelium specific receptor tyrosine kinase 2 antibody
    • hTIE 2 antibody
    • hTIE2 antibody
    • Hyk antibody
    • p140 TEK antibody
    • Soluble TIE2 variant 1 antibody
    • Soluble TIE2 variant 2 antibody
    • Tek antibody
    • tek tyrosine kinase antibody
    • TEK tyrosine kinase endothelial antibody
    • tek tyrosine kinase, endothelial antibody
    • TIE 2 antibody
    • TIE2 antibody
    • TIE2_HUMAN antibody
    • Tunica interna endothelial cell kinase antibody
    • Tyrosine kinase with Ig and EGF homology domains 2 antibody
    • Tyrosine kinase with Ig and EGF homology domains-2 antibody
    • Tyrosine protein kinase receptor TEK antibody
    • Tyrosine protein kinase receptor TIE 2 antibody
    • Tyrosine-protein kinase receptor TEK antibody
    • Tyrosine-protein kinase receptor TIE-2 antibody
    • Venous malformations multiple cutaneous and mucosal antibody
    • VMCM 1 antibody
    • VMCM antibody
    • VMCM1 antibody
    see all

Anti-TIE2 antibody [Cl. 16] images


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 126 kDa


    Exposure time : 2 minutes
  • Overlay histogram showing JEG-3 cells stained with ab24859 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab24859, 2µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in JEG-3 cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

    Please note that Abcam does not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.
  • ab24859 staining TIE2 in Human spleen by Immunohistochemistry (Frozen sections).
  • All lanes : Anti-TIE2 antibody [Cl. 16] (ab24859)

    Lane 1 : HUVECs left untreated
    Lane 2 : HUVECs stimulated for 3 hours with PMA at 25 ng/ml
    Lane 3 : HUVECs stimulated for 6 hours with PMA at 25 ng/ml
    Lane 4 : HUVECs stimulated for 9 hours with PMA at 25 ng/ml
    Lane 5 : HUVECs stimulated for 24 hours with PMA at 25 ng/ml


    Predicted band size : 126 kDa
    Samples were immunoprecipitated with another TIE2 antibody.
  • Immunohistochemical analysis of Human brain tissue, staining TIE2 with ab24859.

    Tissue was fixed with paraformaldehyde, permeabilized with 0.25 Triton X-100 and blocked with 2.5% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/200 in 2.5% horse serum) for 18 hours at 4°C. An HRP-conjugated horse anti-rabbit polyclonal IgG was used as the secondary antibody.

    See Abreview

References for Anti-TIE2 antibody [Cl. 16] (ab24859)

This product has been referenced in:
  • Stiles JM  et al. Targeting of beta adrenergic receptors results in therapeutic efficacy against models of hemangioendothelioma and angiosarcoma. PLoS One 8:e60021 (2013). WB . Read more (PubMed: 23555867) »
  • Soblet J  et al. Variable Somatic TIE2 Mutations in Half of Sporadic Venous Malformations. Mol Syndromol 4:179-83 (2013). WB . Read more (PubMed: 23801934) »

See all 5 Publications for this product

Product Wall

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (Hep G2)
Specification Hep G2
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Jan 08 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/HCl
Sample Mouse Tissue sections (melanoma tumour from C57 mice)
Specification melanoma tumour from C57 mice
Permeabilization Yes - Tween 20 0,5%
Fixative Paraformaldehyde
Username

Mr. Javier Rodriguez

Verified customer

Submitted Nov 14 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Antigen retrieval step None
Sample Human Tissue sections (Arterial wall)
Specification Arterial wall
Permeabilization No
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 25 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Sample Human Tissue sections (Brain)
Specification Brain
Permeabilization No
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 29 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Brain)
Specification Brain
Fixative Paraformaldehyde
Permeabilization Yes - 0.25% Triton X-100
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Oct 26 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Mouse Tissue sections (Liver)
Specification Liver
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization Yes - 0.25% Triton X-100
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5 · Temperature: 25°C
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Verified customer

Submitted Oct 16 2012

Thank you for your patience in this matter and I do apologize for the delay.

Unfortunately we do not have any information about any agonistic activity of our anti-human TIE-2, ab24859.

I am sorry about that I cannot provide any more ...

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Thank you for contacting to Abcam.


Please find the answers to your questions below:

Question 1: I was informed by the lab that epitope mapping was carried out on this antibody a while back, and they are looking through their note...

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Your credit note ID is XXXXXXX.
I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of t...

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We have same lot in stock so would you like to try the same lot? Alternatively I can provide ab71712 which was tested in mouse and is predicted to react with human species. This antibody hasn't ...

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