Recombinant Human Erythropoietin/EPO Protein, CF

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 0.075-0.750 ng/mL.

2 μg/lane of Recombinant Human Erythropoietin/EPO Protein (Catalog # 11264-TC) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing bands at 33-41 kDa.

• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human Erythropoietin/EPO Protein, CF 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 0.075-0.750 ng/mL.
• Source: Chinese Hamster Ovary cell line, CHO-derived human Erythropoietin/EPO protein
  Ala28-Arg193
• Accession #: NP_000790.2
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 21 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: 33-41 kDa, under 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 Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 100-500 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 Erythropoietin/EPO Protein, CF

• 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, 3). Glycosylation of the EPO protein is required for biological activities in vivo (4). The mature human EPO protein shares 75% - 84% amino acid sequence identity with bovine, canine, equine, feline, 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 (5). It is also produced in much lower, but functionally significant amounts by fetal hepatocytes and in adult liver and brain (6-8). EPO promotes erythrocyte formation by preventing the apoptosis of early erythroid precursors which express the erythropoietin receptor (EPO R) (8, 9). EPO R has also been described in brain, retina, heart, skeletal muscle, kidney, endothelial cells, and a variety of tumor cells (7, 8, 10, 11). Ligand induced dimerization of EPO R triggers JAK2-mediated signaling pathways followed by receptor/ligand endocytosis and degradation (1, 12). Rapid regulation of circulating EPO allows tight control of erythrocyte production and hemoglobin concentrations. Anemia or other causes of low tissue oxygen tension induce erythropoietin production by stabilizing the hypoxia-induceable transcription factors HIF-1 alpha and HIF-2 alpha (1, 6). EPO additionally plays a tissue-protective role in ischemia by blocking apoptosis and inducing angiogenesis (7, 8, 13).

1. Koury, M.J. (2005) Exp. Hematol. 33:1263.
2. Jacobs, K. et al. (1985) Nature 313:806.
3. Wen, D. et al. (1993) Blood 82:1507.
4. Tsuda E., et al. (1990) Eur. J. Biochem. 188:405.
5. Lacombe, C. et al. (1988) J. Clin. Invest. 81:620.
6. Eckardt, K.U. and A. Kurtz (2005) Eur. J. Clin. Invest. 35 Suppl. 3:13.
7. Sharples, E.J. et al. (2006) Curr. Opin. Pharmacol. 6:184.
8. Rossert, J. and K. Eckardt (2005) Nephrol. Dial. Transplant 20:1025.
9. Koury, M.J. and M.C. Bondurant (1990) Science 248:378.
10. Acs, G. et al. (2001) Cancer Res. 61:3561.
11. Hardee, M.E. et al. (2006) Clin. Cancer Res. 12:332.
12. Verdier, F. et al. (2000) J. Biol. Chem. 275:18375.
13. Kertesz, N. et al. (2004) Dev. Biol. 276:101.

Bioinformatics

• Uniprot:
  • NP_000790.2(Human)