Recombinant Human Erythropoietin/EPO Protein

Product Discontinued

Recombinant Human Erythropoietin/EPO Protein Summary

• Description: A biologically active protein corresponding to the signal peptide sequence, and the mature Epo sequence of EPO.Source: modified human 293 cells
• Specificity: A DNA sequence encoding the human EPO protein sequence. EPO migrates as a broad band between 25 and 40 kDa in SDS-PAGE due to post-translation modifications, in particular glycosylation. This compares with the unmodified EPO that has a predicted molecular mass of 18.4 kDa. EPO has N-linked and O-linked oligosaccharides. All 3 N-linked sites are verified by peptide mass fingerprinting.
• Preparation Method: A DNA sequence encoding the human EPO protein sequence (containing the signal peptide sequence, and the mature Epo sequence) was expressed in modified human 293 cells.
• Details of Functionality: The ED50 of EPO is typically 0.2 - 0.8 ng/ml as measured in a cell proliferation assay using the human growth factor-dependent TF-1 cell line.
• Protein/Peptide Type: Biologically Active Protein
• Gene: EPO

Applications/Dilutions

• Dilutions:
  • Block/Neutralize
  • Functional
  • Western Blot
• Application Notes: This protein is functionally active and can be used for Blocking and Neutralizing. It can also be used for Western Blot. The EPO separates into a number of isoforms in 2D PAGE due to the presence of post-translational modifications, in particular glycosylation. The pI of the isoforms range between 4.1 and 8.5. This compares with a predicted pI of 7.88 for the unmodified EPO.

Reactivity Notes

This is a Human protein

Packaging, Storage & Formulations

• Storage: Store at -80C. Avoid freeze-thaw cycles.
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute with 0.5 ml sterilized PBS. After reconstitution the buffer solution will contain 1% human serum albumin and 10% trehalose already present in the vial.

Alternate Names for Recombinant Human Erythropoietin/EPO Protein

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

Background

Erythropoietin (EPO) is a hormone produced primarily by the kidney and is the main regulator of red blood cell production. Its major functions are to promote the differentiation and development of red blood cells and to initiate the production of hemoglobin. EPO acts by binding to a specific erythropoietin receptor (EPOR) present on target cells, the red cell precursors in the bone marrow, stimulating them to transform into mature erythrocytes. Human EPO cDNA encodes a 193 amino acid residue precursor protein that is processed to yield a 165 amino acid residue mature protein. EPO contains one O-linked and three Nlinked glycosylation sites. Glycosylation of EPO is required for EPO biological activities in vivo. Recombinant EPO has been approved for the treatment of anemia associated with chronic renal failure, anemia secondary to AZT treatment of AIDS, and anemia associated with cancer. There is recent evidence that recombinant EPO may be useful for cardioprotection and neuroprotection. This is a HCX protein. HCX Expression System Details HCX proteins mimic the proteins in the human body because they are expressed from human, rather than animal, insect or bacterial cells. This process gives them human post-translational modifications. Recombinant DNA techniques allow a human protein with the correct amino acid sequence to be expressed in a non-human cell line. However, non-human cells lack the appropriate cellular machinery, such as specific glycosyltransferases, necessary to produce the correct human post-translational modifications of a protein. An extreme example is seen in E. coli cells, which produce recombinant proteins with no glycosylation, as the above figure illustrates. Rodent and yeast cells are able to glycosylate proteins, but they are still different from glycosylation in human cells. Expression System Resultant Proteins Human (e.g. K562, HEK293) Correct amino acid sequence Human post-translational modifications Rodent (e.g. CHO, NSO) Correct amino acid sequence Some natural glycosylation - not human-like Yeast (e.g. Pichia) Correct amino acid sequence Some natural glycosylation - not human-like E.Coli Correct amino acid sequence No PTMs Although there have been significant attempts to make non-human cell derived cytokines more human-like, there is a growing awareness that in many instances, particularly in therapeutics, cytokines should mimic those found in the body as closely as possible. Purified Symansis EPO consists of 25-55% carbohydrate by weight.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Peptides and proteins are guaranteed for 2 years from date of receipt.

Publications for Erythropoietin/EPO Biologically Active Protein (NBP1-46048)(1)

Publication using NBP1-46048

• Zhang L, Wang Y, Wang J et al. rhEPO/EPO discrimination with ultrasensitive electrochemical biosensor based on sandwich-type nano-Au/ZnO sol-gel/nano-Au signal amplification. Biosens Bioelectron. 2013-06-27 [PMID: 23867351]

Bioinformatics

• Gene Symbol: EPO