Recombinant Canine Erythropoietin/EPO Protein

• Reactivity: Ca
• Applications: Bioactivity

Recombinant Canine Erythropoietin/EPO Protein Summary

• Details of Functionality: Measured in a cell proliferation assay using TF‑1 human erythroleukemic cells. Kitamura, T. et al. (1989) J. Cell Physiol. 140:323. The ED₅₀ for this effect is 0.4-2 ng/mL.
• Source: Mouse myeloma cell line, NS0-derived canine Erythropoietin/EPO protein
  Ala41-Arg206
• Accession #: P33707
• N-terminal Sequence: Ala41
• Protein/Peptide Type: Recombinant Proteins
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 18.4 kDa.
  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: 36-38 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 with BSA as a carrier protein.
• Purity: >90%, by SDS-PAGE under reducing conditions and visualized by silver stain.
• Reconstitution Instructions: Reconstitute at 10 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin.

Notes

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

Alternate Names for Recombinant Canine Erythropoietin/EPO Protein

• ECYT5
• EP
• EPO
• epoetin
• Erythropoietin
• MGC138142
• MVCD2

Background

Erythropoietin (EPO) is a 34 kDa glycoprotein hormone in the type I cytokine family and is related to thrombopoietin (1). Its three N-glycosylation sites, four alpha helices, and N- to C-terminal disulfide bond are conserved across species (2). Glycosylation of EPO is required for biological activities in vivo (3). Mature canine EPO shares 95% amino acid sequence identity with feline EPO and 80%-89% with bovine, equine, human, mouse, ovine, porcine, and rat EPO. EPO is primarily produced in the kidney by a population of fibroblast-like cortical interstitial cells adjacent to the proximal tubules (4). It is also produced in much lower, but functionally significant amounts by fetal hepatocytes and in adult liver and brain (5-7). EPO promotes erythrocyte formation by preventing the apoptosis of early erythroid precursors which express the EPO receptor (EPO R) (7, 8). EPO R has also been described in brain, retina, heart, skeletal muscle, kidney, endothelial cells, and a variety of tumor cells (6, 7, 9, 10). Ligand induced dimerization of EPO R triggers JAK2-mediated signaling pathways followed by receptor/ligand endocytosis and degradation (1, 11). Rapid regulation of circulating EPO allows tight control of erythrocyte production and hemoglobin concentrations. Anemia or other causes of low tissue oxygen tension induce EPO production by stabilizing the hypoxia-induceable transcription factors HIF-1 alpha and HIF-2 alpha (1, 5). EPO additionally plays a tissue-protective role in ischemia by blocking apoptosis and inducing angiogenesis (6, 7, 12).

1. Koury, M.J. (2005) Exp. Hematol. 33:1263.
2. Wen, D. et al. (1993) Blood 82:1507.
3. Tsuda E., et al. (1990) Eur. J. Biochem. 188:405.
4. Lacombe, C. et al. (1988) J. Clin. Invest. 81:620.
5. Eckardt, K. U. and A. Kurtz (2005) Eur. J. Clin. Invest. 35 Suppl. 3:13.
6. Sharples, E. J. et al. (2006) Curr. Opin. Pharmacol. 6:184.
7. Rossert, J. and K. Eckardt (2005) Nephrol. Dial. Transplant 20:1025.
8. Koury, M.J. and M.C. Bondurant (1990) Science 248:378.
9. Acs, G. et al. (2001) Cancer Res. 61:3561.
10. Hardee, M.E. et al. (2006)Clin. Cancer Res. 12:332.
11. Verdier, F. et al. (2000) J. Biol. Chem. 275:18375.
12. Kertesz, N. et al. (2004) Dev. Biol. 276:101.

Bioinformatics

• Uniprot:
  • P33707()