Tie-2 [p Tyr992] Antibody [Unconjugated]

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.

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.

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.

• Reactivity: Hu, Mu
• Applications: WB, IHC, CyTOF-ready, Flow
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: LYOPH

Tie-2 [p Tyr992] Antibody [Unconjugated] Summary

• Immunogen: Phosphopeptide containing human Tie-2 Y992 site
• Modification: p Tyr992
• Specificity: Detects human and mouse Tie-2 when phosphorylated at Y992 in Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: TEK
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Immunohistochemistry 1-15 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Western Blot 0.5 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Tie-2 [p Tyr992] Antibody [Unconjugated]

• 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

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Tie-2 Antibody (AF2720)(21)

Publications using AF2720

• A Lask, B Gutbier, O Kershaw, G Nouailles, AD Gruber, HC Müller-Red, S Chackowicz, DA Hamilton, P Van Slyke, M Witzenrath Adjunctive therapy with the Tie2 agonist Vasculotide reduces pulmonary permeability in Streptococcus pneumoniae infected and mechanically ventilated mice Scientific Reports, 2022;12(1):15531. 2022 [PMID: 36109537] (Western Blot, Mouse)
• YG Choe, JH Yoon, J Joo, B Kim, SP Hong, GY Koh, DS Lee, WY Oh, Y Jeong Pericyte Loss Leads to Capillary Stalling Through Increased Leukocyte-Endothelial Cell Interaction in the Brain Frontiers in Cellular Neuroscience, 2022;16(0):848764. 2022 [PMID: 35360491] (Western Blot, Mouse)
• 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 Oligodendrocyte precursor cell specification is regulated by bidirectional neural progenitor-endothelial cell crosstalk Nature Neuroscience, 2021;0(0):. 2021 [PMID: 33510480] (IHC, Mouse)
• ME Griffin, AW Sorum, GM Miller, WA Goddard, LC Hsieh-Wils Sulfated glycans engage the Ang-Tie pathway to regulate vascular development Nat Chem Biol, 2020;0(0):. 2020 [PMID: 33020664] (Western Blot, Human)
• J Gu, Y Zhang, Z Han, L Gao, J Cui, Y Sun, Y Niu, B You, CP Huang, C Chang, X Wang, S Yeh 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 Cell Death Dis, 2020;11(5):367. 2020 [PMID: 32409702] (Western Blot, Human)
• 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 Preclinical validation of a novel metastasis-inhibiting Tie1 function-blocking antibody EMBO Mol Med, 2020;0(0):e11164. 2020 [PMID: 32302470] (Western Blot, Mouse)
• K Kato, R Diéguez-Hu, DY Park, SP Hong, S Kato-Azuma, S Adams, M Stehling, B Trappmann, JL Wrana, GY Koh, RH Adams Pulmonary pericytes regulate lung morphogenesis Nat Commun, 2018;9(1):2448. 2018 [PMID: 29934496] (Western Blot, Mouse)
• Y Lu, PWL Tai, J Ai, DJ Gessler, Q Su, X Yao, Q Zheng, PD Zamore, X Xu, G Gao Transcriptome Profiling of Neovascularized Corneas Reveals miR-204 as a Multi-target Biotherapy Deliverable by rAAVs Mol Ther Nucleic Acids, 2018;10(0):349-360. 2018 [PMID: 29499946] (Western Blot, Mouse)
• D Ko, S Kim Cooperation between ZEB2 and Sp1 promotes cancer cell survival and angiogenesis during metastasis through induction of survivin and VEGF Oncotarget, 2018;9(1):726-742. 2018 [PMID: 29416649] (Western Blot, Human)
• 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 Tie2 protects the vasculature against thrombus formation in systemic inflammation J. Clin. Invest., 2018;0(0):. 2018 [PMID: 29360642] (Western Blot, Human)
• 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 Impaired angiopoietin/Tie2 signaling compromises Schlemm's canal integrity and induces glaucoma J. Clin. Invest., 2017;0(0):. 2017 [PMID: 28920924] (IHC, Mouse)
• 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 The endothelial transcription factor ERG mediates Angiopoietin-1-dependent control of Notch signalling and vascular stability Nat Commun, 2017;8(0):16002. 2017 [PMID: 28695891] (ICC, Human)
• DOA Alawo, TA Tahir, M Fischer, DG Bates, SR Amirova, NPJ Brindle Regulation of Angiopoietin Signalling by Soluble Tie2 Ectodomain and Engineered Ligand Trap Sci Rep, 2017;7(1):3658. 2017 [PMID: 28623351] (Western Blot, Human)
• T Shlamkovic, L Aharon, WA Barton, N Papo Utilizing combinatorial engineering to develop Tie2 targeting antagonistic angiopoetin-2 ligands as candidates for anti-angiogenesis therapy Oncotarget, 2017;8(20):33571-33585. 2017 [PMID: 28422724] (Western Blot, Human)
• VM Leppänen, P Saharinen, K Alitalo Structural basis of Tie2 activation and Tie2/Tie1 heterodimerization Proc. Natl. Acad. Sci. U.S.A., 2017;0(0):. 2017 [PMID: 28396439] (Western Blot, Human)
• Binyam Bezabeh Insertion of scFv into the hinge domain of full-length IgG1 monoclonal antibody results in tetravalent bispecific molecule with robust properties MAbs, 2016;0(0):0. 2016 [PMID: 27981887] (Western Blot, Human)
• Angiopoetin-2 Signals Do Not Mediate the Hypervascularization of Islets in Type 2 Diabetes PLoS ONE, 2016;11(9):e0161834. 2016 [PMID: 27617438] (Western Blot, Human)
• 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 Endothelial exocytosis of angiopoietin-2 resulting from CCM3 deficiency contributes to cerebral cavernous malformation. Nat Med, 0;22(9):1033-1042. 0 [PMID: 27548575] (Western Blot, Mouse)
• Woo KV, Qu X, Babaev VR, Linton MF, Guzman RJ, Fazio S, Baldwin HS Tie1 attenuation reduces murine atherosclerosis in a dose-dependent and shear stress-specific manner. J. Clin. Invest., 2011;121(4):1624-35. 2011 [PMID: 21383501] (Western Blot, Mouse)
• Bai Y, Cui M, Meng Z, Shen L, He Q, Zhang X, Chen F, Xiao J Ectopic expression of angiopoietin-1 promotes neuronal differentiation in neural progenitor cells through the Akt pathway. Biochem. Biophys. Res. Commun., 2009;378(2):296-301. 2009 [PMID: 19028450] (Western Blot, Mouse)
• Saharinen P, Eklund L, Miettinen J, Wirkkala R, Anisimov A, Winderlich M, Nottebaum A, Vestweber D, Deutsch U, Koh GY, Olsen BR, Alitalo K Angiopoietins assemble distinct Tie2 signalling complexes in endothelial cell-cell and cell-matrix contacts. Nat. Cell Biol., 2008;10(5):527-37. 2008 [PMID: 18425119] (ICC, Human)

Review for Tie-2 Antibody (AF2720) (1)

Average Rating: 3

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Bhavin Shah Immunofluorescence in mouse brain tissue Mouse 10/02/2020

Summary

• Application: Other
• Sample Tested: Cerebellum tissue
• Species: Mouse
• Lot: VRQ0314031

Comments

• Comments: Postnatal mouse cerebellum at P14 was used to determine the levels of phospho-Tie2 in the blood vessels. The blocking buffer and the antibody buffers were prepared in 0.2% gelatin in PBST (In primary and secondary solutions were used 0.025% of Saponin was added). The sections were antigen retrieved by boiling in a microwave for 5 min using 10mM Sodium citrate (pH6).

Bioinformatics

• Gene Symbol: TEK