Human/Mouse Phospho-Tie-2 (Y992) Antibody

Catalog # Availability Size / Price Qty
AF2720
AF2720-SP
Detection of Mouse Phospho-Tie‑2 (Y992) by Western Blot.
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Product Details
Citations (32)
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Human/Mouse Phospho-Tie-2 (Y992) Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects human and mouse Tie-2 when phosphorylated at Y992 in Western blots.
Source
Polyclonal Rabbit IgG
Purification
Antigen Affinity-purified
Immunogen
Phosphopeptide containing human Tie-2 Y992 site
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
0.5 µg/mL
See below
Immunohistochemistry
1-15 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Intracellular Staining by Flow Cytometry
0.25 µg/106 cells
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot Detection of Mouse Phospho-Tie-2 (Y992) antibody by Western Blot. View Larger

Detection of Mouse Phospho-Tie‑2 (Y992) by Western Blot. Western blot shows lysates of NIH-3T3 mouse embryonic fibroblast cell line transfected with mouse Tie-2 and untreated (-) or treated (+) with 600 ng/mL Recombinant Human Angiopoietin-1 (Catalog # 923-AN) for 5 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human/Mouse Phospho-Tie-2 (Y992) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2720) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-Tie-2 (Y992) at approximately 150 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry Phospho-Tie-2 (Y992) antibody in Human Kidney Tissue by Immunohistochemistry (IHC-P). View Larger

Phospho-Tie-2 (Y992) in Human Kidney Tissue. Tie-2 was detected in immersion fixed paraffin-embedded sections of human kidney tissue using Rabbit Anti-Human/Mouse Phospho-Tie-2 (Y992) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2720) at 1 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (brown; Catalog # VC003) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in convoluted tubules. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Intracellular Staining by Flow Cytometry Detection of Tie‑2 antibody in HUVEC Human Cells antibody by Flow Cytometry. View Larger

Detection of Tie‑2 in HUVEC Human Cells by Flow Cytometry. HUVEC human umbilical vein endothelial cells were unstimulated (light orange filled histogram) or treated with 100 µM pervanadate for 15 minutes (dark orange filled histogram), then stained with Human/Mouse Phospho- Tie-2 (Y992) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2720) or control antibody (Catalog # AB-105-C, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.

Western Blot Detection of Mouse Tie-2 by Western Blot View Larger

Detection of Mouse Tie-2 by Western Blot Pericyte-derived Angpt1 controls alveologenesis. a RT-qPCR analysis of Angpt1 and Tie2/Tek expression in freshly sorted lung GFP+, CD31+ or EpCAM+ cells from P7 Pdgfrb(BAC)-CreERT2 R26-mT/mG mice. Data represents mean ± s.e.m. (n = 4 mice). b High magnification images of P10 Angpt1GFP lungs stained for GFP (green), PDGFR beta (red), and PDGFR alpha (blue). Arrows indicate GFP and PDGFR beta double positive pericytes. Scale bar, 15 µm. c RT-qPCR analysis of Angpt1 expression in freshly sorted PDGFR beta + cells from P7 Yap1,Wwtr1iPCKO and control lungs. Data represents mean ± s.e.m. (n = 4 mice, two-tailed unpaired t-test). dAngpt1 expression in cultured Verteporfin (VP)-treated (48 h) and control pericytes. Data represents mean ± s.e.m. (n = 4, Welch’s t-test). e Expression of the indicated transcripts in freshly sorted CD31+ cells from P7 Yap1,Wwtr1iPCKO and control lungs. Data represents mean ± s.e.m. (n = 4 mice, NS not significant, two-tailed unpaired t-test). f–h Western blot analysis of Angpt1 protein (f; n = 2 controls and 4 mutant mice) and of total and phospho-Tie2 (pTie2) in P12 Yap1,Wwtr1iPCKO and control total lung lysates (g, n = 3 controls and 5 mutants). Molecular weight marker (kDa) is indicated. Relative quantification of signals is shown in h. Two-tailed unpaired t-test. i Scheme showing the time points of tamoxifen administration and analysis for Angpt1iPCKO mice. j, k 3D reconstruction confocal images of P12 Angpt1iPCKO and littermate control lungs stained for AQP5 (green), PDGFR beta (red), and PECAM1 (blue). Panels in k show higher magnification of PECAM1 staining. Scale bar, 50 µm (j) and 30 µm (k). l Quantitation of airspace volume in P12 Angpt1iPCKO and littermate control lung sections with 3D reconstruction surface images. Data represents mean ± s.e.m. (n = 4 mice; p < 0.0001, two-tailed unpaired t-test). m 3D reconstruction confocal images of P12 Angpt1iPCKO and littermate control lungs stained for alpha SMA (red) and tropoelastin (blue). Scale bar, 50 µm. n Quantitation of staining intensity for alpha SMA or tropoelastin shown in m. Intensity was normalized to the size of the parenchymal region. Data represents mean ± s.e.m. (n = 4 mice; NS not significant, two-tailed unpaired t-test). o Schematic summary of findings. Pulmonary pericytes regulate ECs and alveolar epithelial cells via angiocrine factors such as angiopoietin-1 and HGF Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/29934496), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Tie-2

Tie-2 (also known as TEK) is an angiogenic receptor tyrosine kinase required for the later stage of blood vessel maturation. Ligand binding induces receptor dimerization and autophosphorylation on multiple tyrosine residues. Y992 is located on the putative activation loop of Tie-2 and is a major autophosphorylation site.

Long Name
Tyrosine Kinase with Immunoglobulin and Epidermal Growth Factor Homology Domains 2
Entrez Gene IDs
7010 (Human); 21687 (Mouse); 89804 (Rat); 396729 (Porcine); 403714 (Canine); 102122204 (Cynomolgus Monkey); 30747 (Zebrafish)
Alternate Names
angiopoietin-1 receptor; CD202b antigen; CD202b; EC 2.7.10; EC 2.7.10.1; hTIE2; p140 TEK; soluble TIE2 variant 1; soluble TIE2 variant 2; TEK tyrosine kinase, endothelial; TEK; Tie2; Tie-2; TIE2CD202b; Tunica interna endothelial cell kinase; Tyrosine-protein kinase receptor TEK; Tyrosine-protein kinase receptor TIE-2; venous malformations, multiple cutaneous and mucosal; VMCM; VMCM1

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Citations for Human/Mouse Phospho-Tie-2 (Y992) Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

32 Citations: Showing 1 - 10
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  1. Targeting the Tie2-?v?3 integrin axis with bi-specific reagents for the inhibition of angiogenesis
    Authors: T Shlamkovic, L Aharon, D Koslawsky, Y Einav, N Papo
    BMC Biol., 2018-08-17;16(1):92.
  2. VE-PTP stabilizes VE-cadherin junctions and the endothelial barrier via a phosphatase-independent mechanism
    Authors: Juettner, VV;Kruse, K;Dan, A;Vu, VH;Khan, Y;Le, J;Leckband, D;Komarova, Y;Malik, AB;
    J. Cell Biol.
  3. An agonistic anti-Tie2 antibody suppresses the normal-to-tumor vascular transition in the glioblastoma invasion zone
    Authors: E Lee, EA Lee, E Kong, H Chon, M Llaiqui-Co, CH Park, BY Park, NR Kang, JS Yoo, HS Lee, HS Kim, SH Park, SW Choi, D Vestweber, JH Lee, P Kim, WS Lee, I Kim
    Experimental & Molecular Medicine, 2023-02-24;55(2):470-484.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  4. Shear stress control of vascular leaks and atheromas through Tie2 activation by VE-PTP sequestration
    Authors: K Shirakura, P Baluk, AF Nottebaum, U Ipe, KG Peters, DM McDonald, D Vestweber
    Embo Molecular Medicine, 2023-02-06;0(0):e16128.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  5. Adjunctive therapy with the Tie2 agonist Vasculotide reduces pulmonary permeability in Streptococcus pneumoniae infected and mechanically ventilated mice
    Authors: A Lask, B Gutbier, O Kershaw, G Nouailles, AD Gruber, HC Müller-Red, S Chackowicz, DA Hamilton, P Van Slyke, M Witzenrath
    Scientific Reports, 2022-09-15;12(1):15531.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  6. Pericyte Loss Leads to Capillary Stalling Through Increased Leukocyte-Endothelial Cell Interaction in the Brain
    Authors: YG Choe, JH Yoon, J Joo, B Kim, SP Hong, GY Koh, DS Lee, WY Oh, Y Jeong
    Frontiers in Cellular Neuroscience, 2022-03-11;16(0):848764.
    Species: Mouse
    Sample Types: Protein Lysates
    Applications: Western Blot
  7. Oligodendrocyte precursor cell specification is regulated by bidirectional neural progenitor-endothelial cell crosstalk
    Authors: I Paredes, JR Vieira, B Shah, CF Ramunno, J Dyckow, H Adler, M Richter, G Schermann, E Giannakour, L Schirmer, HG Augustin, C Ruiz de Al
    Nature Neuroscience, 2021-01-28;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  8. Sulfated glycans engage the Ang-Tie pathway to regulate vascular development
    Authors: ME Griffin, AW Sorum, GM Miller, WA Goddard, LC Hsieh-Wils
    Nat Chem Biol, 2020-10-05;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  9. Targeting the ER&beta/Angiopoietin-2/Tie-2 signaling-mediated angiogenesis with the FDA-approved anti-estrogen Faslodex to increase the Sunitinib sensitivity in RCC
    Authors: J Gu, Y Zhang, Z Han, L Gao, J Cui, Y Sun, Y Niu, B You, CP Huang, C Chang, X Wang, S Yeh
    Cell Death Dis, 2020-05-14;11(5):367.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  10. Preclinical validation of a novel metastasis-inhibiting Tie1 function-blocking antibody
    Authors: M Singhal, N Gengenbach, S La Porta, S Gehrs, J Shi, M Kamiyama, DM Bodenmille, A Fischl, B Schieb, E Besemfelde, S Chintharla, HG Augustin
    EMBO Mol Med, 2020-04-17;0(0):e11164.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  11. miR-15a/-16 Inhibit Angiogenesis by Targeting the Tie2 Coding Sequence: Therapeutic Potential of a miR-15a/16 Decoy System in Limb Ischemia
    Authors: Marie Besnier, Saran Shantikumar, Maryam Anwar, Parul Dixit, Aranzazu Chamorro-Jorganes, Walid Sweaad et al.
    Molecular Therapy - Nucleic Acids
  12. MicroRNA-711–Induced Downregulation of Angiopoietin-1 Mediates Neuronal Cell Death
    Authors: Boris Sabirzhanov, Alan I. Faden, Taryn Aubrecht, Rebecca Henry, Ethan Glaser, Bogdan A. Stoica
    Journal of Neurotrauma
  13. Integrin alpha 5 beta 1-Ang1/Tie2 receptor cross-talk regulates brain endothelial cell responses following cerebral ischemia
    Authors: Defang Pang, Lu Wang, Jing Dong, Xiaoyin Lai, Qijuan Huang, Richard Milner et al.
    Experimental & Molecular Medicine
  14. Pulmonary pericytes regulate lung morphogenesis
    Authors: K Kato, R Diéguez-Hu, DY Park, SP Hong, S Kato-Azuma, S Adams, M Stehling, B Trappmann, JL Wrana, GY Koh, RH Adams
    Nat Commun, 2018-06-22;9(1):2448.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  15. Modulation of Angiopoietin 2 release from endothelial cells and angiogenesis by the synaptic protein Neuroligin 2
    Authors: Margherita Pergolizzi, Laura Bizzozero, Elena Riccitelli, Davide Pascal, Anna Valeria Samarelli, Federico Bussolino et al.
    Biochemical and Biophysical Research Communications
  16. Tie2 protects the vasculature against thrombus formation in systemic inflammation
    Authors: SJ Higgins, K Ceunynck, J Kellum, X Chen, X Gu, SA Chaudhry, S Schulman, TA Libermann, S Lu, NI Shapiro, DC Christiani, R Flaumenhaf, SM Parikh
    J. Clin. Invest., 2018-03-05;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  17. Transcriptome Profiling of Neovascularized Corneas Reveals miR-204 as a Multi-target Biotherapy Deliverable by rAAVs
    Authors: Y Lu, PWL Tai, J Ai, DJ Gessler, Q Su, X Yao, Q Zheng, PD Zamore, X Xu, G Gao
    Mol Ther Nucleic Acids, 2017-12-30;10(0):349-360.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  18. Cooperation between ZEB2 and Sp1 promotes cancer cell survival and angiogenesis during metastasis through induction of survivin and VEGF
    Authors: D Ko, S Kim
    Oncotarget, 2017-12-11;9(1):726-742.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  19. Impaired angiopoietin/Tie2 signaling compromises Schlemm's canal integrity and induces glaucoma
    Authors: J Kim, DY Park, H Bae, DY Park, D Kim, CK Lee, S Song, TY Chung, DH Lim, Y Kubota, YK Hong, Y He, HG Augustin, G Oliver, GY Koh
    J. Clin. Invest., 2017-09-18;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  20. The endothelial transcription factor ERG mediates Angiopoietin-1-dependent control of Notch signalling and vascular stability
    Authors: AV Shah, GM Birdsey, C Peghaire, ME Pitulescu, NP Dufton, Y Yang, I Weinberg, L Osuna Alma, L Payne, JC Mason, H Gerhardt, RH Adams, AM Randi
    Nat Commun, 2017-07-11;8(0):16002.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  21. Regulation of Angiopoietin Signalling by Soluble Tie2 Ectodomain and Engineered Ligand Trap
    Authors: DOA Alawo, TA Tahir, M Fischer, DG Bates, SR Amirova, NPJ Brindle
    Sci Rep, 2017-06-16;7(1):3658.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  22. Utilizing combinatorial engineering to develop Tie2 targeting antagonistic angiopoetin-2 ligands as candidates for anti-angiogenesis therapy
    Authors: T Shlamkovic, L Aharon, WA Barton, N Papo
    Oncotarget, 2017-05-16;8(20):33571-33585.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  23. Structural basis of Tie2 activation and Tie2/Tie1 heterodimerization
    Authors: VM Leppänen, P Saharinen, K Alitalo
    Proc. Natl. Acad. Sci. U.S.A., 2017-04-10;0(0):.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: ICC, Western Blot
  24. Insertion of scFv into the hinge domain of full-length IgG1 monoclonal antibody results in tetravalent bispecific molecule with robust properties
    Authors: Binyam Bezabeh
    MAbs, 2017-02-01;0(0):0.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  25. Angiopoetin-2 Signals Do Not Mediate the Hypervascularization of Islets in Type 2 Diabetes
    PLoS ONE, 2016-09-12;11(9):e0161834.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  26. Endothelial exocytosis of angiopoietin-2 resulting from CCM3 deficiency contributes to cerebral cavernous malformation.
    Authors: Jenny Zhou H, Qin L, Zhang H, Tang W, Ji W, He Y, Liang X, Wang Z, Yuan Q, Vortmeyer A, Toomre D, Fuh G, Yan M, Kluger M, Wu D, Min W
    Nat Med, 2016-08-22;22(9):1033-1042.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  27. Effects of Angiopoietin-1 on Hemorrhagic Transformation and Cerebral Edema after Tissue Plasminogen Activator Treatment for Ischemic Stroke in Rats
    Authors: Kunio Kawamura, Tetsuya Takahashi, Masato Kanazawa, Hironaka Igarashi, Tsutomu Nakada, Masatoyo Nishizawa et al.
    PLoS ONE
  28. Neurovascular Signals Suggest a Propagation Mechanism for Endogenous Stem Cell Activation Along Blood Vessels
    Authors: Jimmy Masjkur, Maria Adele Adele Rueger, Stefan R R. Bornstein, Ron McKay, Andreas Androutsellis-Theotokis
    CNS & Neurological Disorders - Drug Targets
  29. Tie1 attenuation reduces murine atherosclerosis in a dose-dependent and shear stress-specific manner.
    Authors: Woo KV, Qu X, Babaev VR, Linton MF, Guzman RJ, Fazio S, Baldwin HS
    J. Clin. Invest., 2011-03-07;121(4):1624-35.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  30. Cholera Toxin Regulates a Signaling Pathway Critical for the Expansion of Neural Stem Cell Cultures from the Fetal and Adult Rodent Brains
    Authors: Andreas Androutsellis-Theotokis, Stuart Walbridge, Deric M. Park, Russell R. Lonser, Ronald D. G. McKay
    PLoS ONE
  31. Ectopic expression of angiopoietin-1 promotes neuronal differentiation in neural progenitor cells through the Akt pathway.
    Authors: Bai Y, Cui M, Meng Z, Shen L, He Q, Zhang X, Chen F, Xiao J
    Biochem. Biophys. Res. Commun., 2008-11-24;378(2):296-301.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  32. Angiopoietins assemble distinct Tie2 signalling complexes in endothelial cell-cell and cell-matrix contacts.
    Authors: Saharinen P, Eklund L, Miettinen J, Wirkkala R, Anisimov A, Winderlich M, Nottebaum A, Vestweber D, Deutsch U, Koh GY, Olsen BR, Alitalo K
    Nat. Cell Biol., 2008-04-20;10(5):527-37.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC

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Human/Mouse Phospho-Tie-2 (Y992) Antibody
By Mohammad Daud Khan on 10/17/2018
Application: WB Sample Tested: HASMC human aortic smooth muscle cells Species: Human