Recombinant Human Angiostatin Fc Chimera Protein, CF

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Angiostatin Fc Chimera Protein, CF Summary

• Details of Functionality: Measured by the ability of the immobilized protein to support the adhesion of THP‑1 human acute monocytic leukemia cells. Chavakis, T. et al. (2005) Blood 105:1036. The ED₅₀ for this effect is 0.5-2.5 μg/mL.
• Source: Mouse myeloma cell line, NS0-derived human Angiostatin protein
  

  Met-Asp	Human IgG1 (Pro100-Lys330)	IEGR	Human Angiostatin (Kringle 1-4) (Val98-Ala459) Accession # P00747
  N-terminus			C-terminus

• Accession #: P00747
• N-terminal Sequence: Met-Asp-Pro₁₀₀
• Structure / Form: Disulfide-linked homodimer
• Protein/Peptide Type: Recombinant Proteins
• Gene: PLG
• Purity: >95%, by SDS-PAGE with silver staining, under reducing conditions
• Endotoxin Note: <0.01 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 67.7 kDa (monomer).
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 75-85 kDa, reducing conditions

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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >95%, by SDS-PAGE with silver staining, under reducing conditions
• Reconstitution Instructions: Reconstitute at 200 μg/mL in PBS.

Notes

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

Alternate Names for Recombinant Human Angiostatin Fc Chimera Protein, CF

• Angiostatin
• HAE4

Background

Angiostatin is an anti‑angiogenic 38‑45 kDa proteolytic fragment of Plasminogen, a 92‑100 kDa glycosylated blood zymogen that serves as the precursor for Plasmin (1). Plasminogen is produced primarily in the liver, but also in other tissues (2). Angiostatin circulates in the plasma, binds endothelial cells and myeloid cells, is present in platelet granules, and is excreted in the urine (1, 3, 9, 10). Human Plasminogen contains an N‑terminal activation peptide between amino acids (aa) 1‑98, five characteristically folded kringle domains (aa 103‑561), and a peptidase S1 domain (aa 581‑808). Cleavage of the activation peptide produces mature Plasminogen, while further cleavage between Arg580 and Val581 by tPA (tissue plasminogen activator) produces the disulfide‑linked two‑subunit enzyme plasmin that dissolves fibrin clots (1, 3). Angiostatin was first identified as consisting of kringles 1‑4, a form called K1‑4 (1, 3, 4). Human Angiostatin (K1‑4, aa 99‑459) shares 79‑80% aa sequence identity with mouse, rat, canine, feline, porcine and bovine K1‑4. Other anti‑angiogenic forms include kringles 1‑3 (K1‑3), or 1‑4 plus most of kringle 5 (K4.5) (3, 4). K4.5, which is reported to be the most active form, occurs in vivo by autoproteolysis of mature Plasminogen in the presence of either a sulfhydryl donor or cell surface actin, while matrix metalloproteins such as MMP3, 7, 9 and 19 can create multiple forms (5‑8). Some primary tumors promote production of Angiostatin, which paradoxically inhibits their metastatic growth (1). Angiostatin has documented anti‑angiogenic and antitumor activity (3). It interferes with FGF‑2 and VEGF signaling, and inhibits endothelial cell proliferation and tube formation, (1, 3, 4, 8). It also inhibits endothelial cell and macrophage migration, and up‑regulates expression of anti‑angiogenic thrombospondin‑1 and macrophage IL‑12 (3, 11‑13). It promotes endothelial cell apoptosis by down‑regulating Bcl‑2 (3, 4, 12).

1. O’Reilly, M.S. et al. (1994) Cell 79:315.
2. Zhang, L. et al. (2002) Thromb. Haemost. 87:493.
3. Soff, G.A. (2000) Cancer Metastasis Rev. 19:97.
4. Chen, Y.H. et al. (2006) Thromb. Haemost. 95:668.
5. Wang, H. et al. (2004) Cancer Res. 64:162.
6. Lijnen, H.R. et al. (1998) Biochemistry 37:4699.
7. Patterson, B.C. and Q.A. Sang (1997) J. Biol. Chem. 272:28823.
8. Brauer, R. et al. (2011) BMC Biochem. 12:38.
9. Radziwon-Balicka, A. et al. (2013) PLoS ONE 8:e59281.
10. Chavakis, T.et al. (2005) Blood 105:1036.
11. Perri, S.R. et al. (2007) FASEB J. 21:3928.
12. Lee, T.Y. et al. (2009) Blood 114:1987.
13. Albini, A. et al. (2009) J. Transl. Med. 7:5.

Bioinformatics

• Gene Symbol: PLG
• Uniprot:
  • P00747()