Erythropoietin, Recombinant, Human (EPO) (Human 293 Cells)
|Molecular Biology||Storage: -20°CShipping: Blue Ice|
Human Cell Expression Cytokines:
Expression of proteins in our Human 293-cell expression system is a cost-effective and scalable production method for high-authenticity, recombinant human proteins.
Functional activities of human proteins depend on proper folding, phosphorylation, disulfide bridge formation, and proteolytic processing or appropriate glycosylation. Ideally, recombinant human proteins are produced in human cells that are more authentic in terms of both physical properties and biochemical functions. However, the current process of human cell expression requires a large quantity of DNA and medium supplemented with bovine serum. Hence, it is often a daunting task to produce sufficient amount of high quality proteins for drug discovery or diagnostic development (1-100mg) at acceptable cost. United States Biological is now supplying a high yield, cost-effective and versatile system to produce active recombinant human proteins in a human cell line to meet the special needs of the broad scientific community and drug discovery industry. In this system, we have broken key bottleneck steps to increase protein production and reduce cost, including
• Engineered human cell line adapted to suspension for growth in serum-free and chemically defined medium
• Proprietary expression vectors driven by strong promoters and signal peptides
• Large scale transient expression with small quantity of DNA
• Rapid creation of stable human cell lines and adaptation to serum-free medium
• Short time to scale up production
• Efficient tag or tag-free purification
Using this expression system, we have produced a large number of highly active human cytokines, kinases and phosphatases. Our technology is particularly suitable for the production of high-authenticity human proteins in medium scale as research and diagnostic reagents, drug screening targets, and therapeutic proteins for preclinical evaluation. Key differences of recombinant human protein production in different systems are illustrated below:
Expression system E. coli Insect cell CHO cell Human cell
Protein folding + ++ +++ ++++
Phosphorylation - ++ +++ ++++
Proteolytic processing - + +++ ++++
Glycosylation - Poor Not human-like Authentic
Human erythropoietin (EPO) is an acidic glycoprotein. It is mainly produced by the kidney. As the primary regulator of the production of red cells, its major functions are to promote erythroid progenitor cells differentiation and to start the synthesis of hemoglobin. EPO acts by binding to a specific erythropoietin receptor (EPOR) on target cells; the red cell precursors in the bone
marrow, and induces their transformation into mature erythrocytes. The EPO sensitivity increases with differentiation of immature progenitor cells. EPO may act as a neuron protector against glutamate toxicity. Its neuroprotective activity comes into effect by neutralizing the toxicity of free radicals. EPO is produced in brain after oxidative stress. EPO is upregulated by hypoxia and prevents apoptosis of erythroid progenitors in bone marrow. EPO has also angiogenic properties. Erythropoietin (EPO) is a glycoprotein hormone produced primarily by the kidney and is the mainfactor regulating red blood cell production. The mature protein consists of a 165 amino acid
polypeptide chain heavily glycosylated at three N-linked and O-linked glycosylation sites yielding atotal molecular mass of 35-45kD. About 40% of the fully glycosylated EPO molecule consists of carbohydrate.
The specific activity was determined by the dose-dependent stimulation of the proliferation of human TF-1 cells (human erythroleukemic indicator cell line).
Endotoxin: 1EU/ug (LAL)
Storage and Stability:
Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile PBS, 0.1% endotoxin-free recombinant HSA.. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Source: HEK 293 cells
Purity: ~ 95% as determined by SDS-PAGE and stained with Coomassie blue
Form: Supplied as a lyophilized powder from 10mM MES pH 6.0, 80mM sodium chloride. Endotoxin free (<1EU/ug).
Reconstitute with sterile PBS, 0.1% endotoxin-free recombinant HSA.
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.