Angiopoietin-2 Antibody (85816) [Unconjugated]

Angiopoietin‑2 was detected in immersion fixed paraffin-embedded sections of human hepatocellular carcinoma tissue (right panel; positive stain) using Mouse Anti-Human/Mouse Angiopoietin‑2 Monoclonal Antibody (Catalog # MAB098) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). 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 DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to secreted and plasma membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Angiopoietin‑2 was detected in perfusion fixed frozen sections of mouse lymph node tissue using Mouse Anti-Human/Mouse Angiopoietin‑2 Monoclonal Antibody (Catalog # MAB098) 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-Mouse IgG VisUCyte™ HRP Polymer Antibody (brown; Catalog # VC001) and counterstained with hematoxylin (blue). Specific staining was localized to cell membrane. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Angiopoietin‑2 was detected in perfusion fixed frozen sections of mouse colon tissue using Mouse Anti-Human/Mouse Angiopoietin‑2 Monoclonal Antibody (Catalog # MAB098) 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-Mouse IgG VisUCyte™ HRP Polymer Antibody (brown; Catalog # VC001) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes in colon mucosa. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

• Reactivity: Hu, Mu
• Applications: IHC, ELISA(Cap)
• Clone: 85816
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated

Angiopoietin-2 Antibody (85816) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human Angiopoietin-2
  Asp68-Phe496
  Accession # O15123
• Specificity: Detects human and mouse Angiopoietin-2 in ELISAs. In ELISAs, no cross-reactivity or interference with recombinant human (rh) Ang‑1, rhAng-4, rhAng-X, recombinant mouse (rm) Ang-3, rmANGPTL-3, rhTie-1, rhTie-2, and rmTie-2 is observed.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: ANGPT2
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • ELISA Capture (Matched Antibody Pair) 2-8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.5-2.0 ug/mL
  • ELISA Standard (Matched Pair)
  • Immunohistochemistry 1-25 ug/mL
• Application Notes: ELISA Detection: Human Angiopoietin-2 Biotinylated Antibody (Catalog number BAM0981)
  Standard: Recombinant Human Angiopoietin-2 (Catalog number 623-AN)

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
• Reconstitution Instructions: Reconstitute at 0.5 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 Angiopoietin-2 Antibody (85816) [Unconjugated]

• AGPT2
• ANG2
• ANG-2
• angiopoietin 2
• Angiopoietin-2
• angiopoietin-2a
• angiopoietin-2B
• angiopoitin 2
• ANGPT2
• Tie2-ligand

Background

Angiopoietin-2 (Ang-2; also ANGPT2) is a secreted glycoprotein that plays a complex role in angiogenesis and inflammation (1, 2). Mature Ang-2 is 478 amino acids (aa) in length. It contains one coiled-coil domain (aa 166 - 248) that mediates multimerization, and a C-terminal fibrinogen-like domain (aa 275 - 495) that mediates receptor binding. Under reducing conditions, secreted monomeric Ang-2 is 65 - 66 kDa in size. Under nonreducing conditions, both natural and recombinant Ang-2 form 140 kDa dimers, 200 kDa trimers, and 250 - 300 kDa tetramers and pentamers (3 - 6). Alternate splicing generates a short isoform that lacks 52 amino acids (aa) preceding the coiled-coil domain (4). Mature human Ang-2 shares 86% aa sequence identity with mouse and rat Ang-2. Ang-2 is widely expressed during development, but it is restricted postnatally to highly angiogenic tissues such as the placenta, ovaries, and uterus (3). It is particularly abundant in vascular endothelial cells (EC) where it is stored in intracellular Weibel-Palade bodies (1, 3, 7). Both Ang-2 and the related Angiopoietin-1 (Ang-1) are ligands for the receptor tyrosine kinase Tie-2 (2). While Ang-1 is a potent Tie-2 agonist, Ang-2 may act as either a Tie-2 antagonist or agonist, depending upon its state of multimerization. The higher the order of oligomer, the more effective Ang-2 becomes as a Tie-2 agonist (3, 8 - 11). The short isoform appears to block the binding of either Ang-1 or full-length Ang-2 to Tie-2 (4). Ang-2 functions as a pro-angiogenic factor, although it can also induce EC death and vessel regression (12, 13). Upon its release from quiescent EC, it regulates vascular remodeling by promoting EC survival, proliferation, and migration and destabilizing the interaction between EC and perivascular cells (8, 13, 14). Ang-2 is required for postnatal vascular remodeling, and it cooperates with Ang-1 during lymphatic vessel development (7, 15). It mediates the upregulation of ICAM-1 and VCAM-1 on EC, which facilitates the adhesion of leukocytes during inflammation (16). Ang-2 is upregulated in both the endothelium and tumor cells of several cancers as well as in ischemic tissue (17 - 20). Its direct interaction with Integrins promotes tumor cell invasion (21, 22). Ang-2 also promotes the neuronal differentiation and migration of subventricular zone progenitor cells (20).

1. Augustin, H.G. et al. (2009) Nat. Rev. Mol. Cell Biol. 10:165.
2. Murdoch, C. et al. (2007) J. Immunol. 178:7405.
3. Maisonpierre, P.C. et al. (1997) Science 27:55.
4. Kim, I. et al. (2000) J. Biol. Chem. 275:18550.
5. Procopio, W.N. et al. (1999) J. Biol. Chem. 274:30196.
6. Kim, K-T. et al. (2005) J. Biol. Chem. 280:20126.
7. Gale, N.W. et al. (2002) Dev. Cell 3:411.
8. Yuan, H.T. et al. (2009) Mol. Cell. Biol. 29:2011.
9. Falcon, B.L. et al. (2009) Am. J. Pathol. 175:2159.
10. Kim, H-Z. et al. (2009) Biochim. Biophys. Acta 1793:772.
11. Kim, I. et al. (2001) Cardiovasc. Res. 49:872.
12. Lobov, I.B. et al. (2002) Proc. Natl. Acad. Sci. 99:11205.
13. Cao, Y. et al. (2007) Cancer Res. 67:3835.
14. Nasarre, P. et al. (2009) Cancer Res. 69:1324.
15. Dellinger, M. et al. (2008) Dev. Biol. 319:309.
16. Fiedler, U. et al. (2006) Nat. Med. 12:235.
17. Koga, K. et al. (2001) Cancer Res. 61:6248.
18. Etoh, T. et al. (2001) Cancer Res. 61:2145.
19. Tressel, S.L. et al. (2008) Arterioscler. Thromb. Vasc. Biol. 28:1989.
20. Liu, X.S. et al. (2009) J. Biol. Chem. 284:22680.
21. Hu, B. et al. (2006) Cancer Res. 66:775.
22. Imanishi, Y. et al. (2007) Cancer Res. 67:4254.

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 Angiopoietin-2 Antibody (MAB098)(6)

Publications using MAB098

• Pezzotta, A;Perico, L;Corna, D;Morigi, M;Remuzzi, G;Benigni, A;Imberti, B; Sirt3 deficiency promotes endothelial dysfunction and aggravates renal injury PloS one 2023-10-10 [PMID: 37816025] (Western Blot, Mouse)
• HJ Zhou, L Qin, Q Jiang, KN Murray, H Zhang, B Li, Q Lin, M Graham, X Liu, J Grutzendle, W Min Caveolae-mediated Tie2 signaling contributes to CCM pathogenesis in a brain endothelial cell-specific Pdcd10-deficient mouse model Nature Communications, 2021-01-25;12(1):504. 2021-01-25 [PMID: 33495460] (ELISA Capture, Human)
• McDonald G, Tabellini L, Storer B, Martin P, Lawler R, Rosinski S, Schoch H, Hansen J Predictive Value of Clinical Findings and Plasma Biomarkers after Fourteen Days of Prednisone Treatment for Acute Graft-versus-host Disease. Biol Blood Marrow Transplant, 2017-05-03;23(8):1257-1263. 2017-05-03 [PMID: 28478120] (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, 2016-08-22;22(9):1033-1042. 2016-08-22 [PMID: 27548575] (Human)
• Vart RJ, Nikitenko LL, Lagos D, Trotter MW, Cannon M, Bourboulia D, Gratrix F, Takeuchi Y, Boshoff C Kaposi's sarcoma-associated herpesvirus-encoded interleukin-6 and G-protein-coupled receptor regulate angiopoietin-2 expression in lymphatic endothelial cells. Cancer Res., 2007-05-01;67(9):4042-51. 2007-05-01 [PMID: 17483315] (ICC, IHC-P, Human)
• Bhandari V, Choo-Wing R, Lee CG, Zhu Z, Nedrelow JH, Chupp GL, Zhang X, Matthay MA, Ware LB, Homer RJ, Lee PJ, Geick A, de Fougerolles AR, Elias JA Hyperoxia causes angiopoietin 2-mediated acute lung injury and necrotic cell death. Nat. Med., 2006-11-05;12(11):1286-93. 2006-11-05 [PMID: 17086189] (IHC-P, Human)

Bioinformatics

• Gene Symbol: ANGPT2
• Uniprot:
  • O15123()