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    tie2-antibody-cl-16-ab24859.pdf

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Signal Transduction Protein Phosphorylation Tyrosine Kinases Receptor Tyrosine Kinases
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Anti-TIE2 antibody [Cl. 16] (ab24859)

  • Datasheet
  • SDS
Reviews (12)Q&A (9)References (11)

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Western blot - Anti-TIE2 antibody [Cl. 16] (ab24859)
  • Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] (ab24859)
  • Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] (ab24859)
  • Flow Cytometry - Anti-TIE2 antibody [Cl. 16] (ab24859)
  • Western blot - Anti-TIE2 antibody [Cl. 16] (ab24859)

Key features and details

  • Mouse monoclonal [Cl. 16] to TIE2
  • Suitable for: IHC-Fr, WB, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-TIE2 antibody [Cl. 16]
    See all TIE2 primary antibodies
  • Description

    Mouse monoclonal [Cl. 16] to TIE2
  • Host species

    Mouse
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-Fr
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

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

  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    Cl. 16
  • Isotype

    IgG1
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Receptor Tyrosine Kinases
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Growth factor receptors
    • Cardiovascular
    • Vasculature
    • Endothelium
    • Stem Cells
    • Endothelial Progenitors
    • Endothelial Markers
    • Developmental Biology
    • Organogenesis
    • Angiogenesis and vasculogenesis
    • Cardiovascular
    • Cardiovascular Markers
    • Cell Markers
    • Endothelial Cells
    • Cardiovascular
    • Angiogenesis
    • Endothelial Cell Markers

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant human TIE2 protein (ab129039)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
IHC-Fr
Human
WB
Human
Application Abreviews Notes
IHC-Fr (2)
Use at an assay dependent concentration.
WB (1)
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 126 kDa.
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.

Notes
IHC-Fr
Use at an assay dependent concentration.
WB
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 126 kDa.
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.

Target

  • Function

    Tyrosine-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 specificity

    Detected 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 disease

    Dominantly inherited venous malformations
    May play a role in a range of diseases with a vascular component, including neovascularization of tumors, psoriasis and inflammation.
  • Sequence similarities

    Belongs 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.
  • Domain

    The 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 localization

    Cell 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.
  • Target information above from: UniProt accession Q02763 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7010 Human
    • Omim: 600221 Human
    • SwissProt: Q02763 Human
    • Unigene: 89640 Human
    • 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

    Images

    • Western blot - Anti-TIE2 antibody [Cl. 16] (ab24859)
      Western blot - Anti-TIE2 antibody [Cl. 16] (ab24859)
    • Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] (ab24859)
      Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] (ab24859)
      ab24859 staining TIE2 in Human spleen by Immunohistochemistry (Frozen sections).
    • Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] (ab24859)
      Immunohistochemistry (Frozen sections) - Anti-TIE2 antibody [Cl. 16] (ab24859)This image is courtesy of an anonymous Abreview
      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

    • Flow Cytometry - Anti-TIE2 antibody [Cl. 16] (ab24859)
      Flow Cytometry - Anti-TIE2 antibody [Cl. 16] (ab24859)
      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.
    • Western blot - Anti-TIE2 antibody [Cl. 16] (ab24859)
      Western blot - Anti-TIE2 antibody [Cl. 16] (ab24859)
      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.

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (11)

    Publishing research using ab24859? Please let us know so that we can cite the reference in this datasheet.

    ab24859 has been referenced in 11 publications.

    • Besnier M  et al. miR-15a/-16 Inhibit Angiogenesis by Targeting the Tie2 Coding Sequence: Therapeutic Potential of a miR-15a/16 Decoy System in Limb Ischemia. Mol Ther Nucleic Acids 17:49-62 (2019). PubMed: 31220779
    • Pang D  et al. Integrin a5ß1-Ang1/Tie2 receptor cross-talk regulates brain endothelial cell responses following cerebral ischemia. Exp Mol Med 50:117 (2018). PubMed: 30185785
    • Yang WJ  et al. Overexpression of Tie2 is associated with poor prognosis in patients with gastric cancer. Oncol Lett 15:8027-8033 (2018). PubMed: 29849805
    • Wang K  et al. The role of angiopoietin-2 in nucleus pulposus cells during human intervertebral disc degeneration. Lab Invest 97:971-982 (2017). PubMed: 28394321
    • Syed M  et al. Hyperoxia causes miR-34a-mediated injury via angiopoietin-1 in neonatal lungs. Nat Commun 8:1173 (2017). PubMed: 29079808
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 21 Abreviews or Q&A

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-TIE2 antibody [Cl. 16]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Rat Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 06 2020

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-TIE2 antibody [Cl. 16]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney
    Blocking step
    Sea Block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 06 2020

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-TIE2 antibody [Cl. 16]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Kidney)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: ab64236 100x Citrate Buffer pH 6.0
    Permeabilization
    Yes - 0.05% Tween 20
    Specification
    Kidney
    Blocking step
    sea block as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More

    Mr. David Ivancic

    Verified customer

    Submitted Dec 24 2019

    IHC - Wholemount abreview for Anti-TIE2 antibody [Cl. 16]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Mouse Tissue (stria)
    Specification
    stria
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 02 2018

    Western blot abreview for Anti-TIE2 antibody [Cl. 16]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Jan 08 2015

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-TIE2 antibody [Cl. 16]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Mr. Javier Rodriguez

    Verified customer

    Submitted Nov 14 2013

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-TIE2 antibody [Cl. 16]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 25 2013

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-TIE2 antibody [Cl. 16]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 29 2013

    Immunohistochemistry (Frozen sections) abreview for Anti-TIE2 antibody [Cl. 16]

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 26 2012

    Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-TIE2 antibody [Cl. 16]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 16 2012

    1-10 of 21 Abreviews or Q&A

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