Active Erythropoietin (EPO)

EP; Epoetin; Erythropoetin; Hematopoietin; Hemopoietin

ACTIVITY TEST

Figure. The binding activity of EPO with EPOR.
Erythropoietin (EPO), also known as hematopoietin or hemopoietin, is a glycoprotein cytokine secreted by the kidney in response to cellular hypoxia. Erythropoietin is an essential hormone for red blood cell production. Its primary effect on red blood cell progenitors and precursors (which are found in the bone marrow in humans) by promoting their survival through protecting these cells from apoptosis, or cell death. EPO is the primary erythropoietic factor that cooperates with various other growth factors involved in the development of erythroid lineage from multipotent progenitors. Besides, Erythropoietin Receptor (EPOR) has been identified as an interactor of EPO, thus a binding ELISA assay was conducted to detect the interaction of recombinant human EPO and recombinant human EPOR. Briefly, EPO were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to EPOR-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-EPO pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of EPO and EPOR was shown in Figure 1, and this effect was in a dose dependent manner.

Figure. Cell proliferation of TF-1 cells after stimulated with EPO.<br/>To test the effect of EPO on cell proliferation, TF-1 cells were seeded into triplicate wells of 96-well plates at a density of 5,000 cells/well with 1% serum standard 1640 including various concentrations of recombinant human EPO. After incubated for 72h, cells were observed by inverted microscope and cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Briefly, 10µL of CCK-8 solution was added to each well of the plate, then the absorbance at 450nm was measured using a microplate reader after incubating the plate for 1-4 hours at 37℃. Proliferation of TF-1 cells after incubation with EPO for 72h observed by inverted microscope was shown in Figure 1. Cell viability was assessed by CCK-8 (Cell Counting Kit-8 ) assay after incubation with recombinant EPO for 72h. The result was shown in Figure 2. It was obvious that EPO significantly increased cell viability of TF-1 cells.<br/>(A) TF-1 cells cultured in 1640, stimulated with 50ng/mL EPO for 72h; <br/>(B) Unstimulated TF-1 cells cultured in1640 for 72h.

Figure. Cell proliferation of TF-1 cells after stimulated with EPO.

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Endocrinology Prenatal Glucocorticoid Overexposure Causes Permanent Increases in Renal Erythropoietin Expression and Red Blood Cell Mass in the Rat Offspring Endo: source
Journal of Ethnopharmacology Inhibitory effect of polysaccharides isolated from Angelica sinensis on hepcidin expression PubMed: 21333724
PLOS ONE CD98 Positive Eosinophils Contribute to T Helper 1 Pattern Inflammation PlosOne: Source
The Journal of Clinical Endocrinology & Metabolism Alterations of Circulating Endothelial Cell and Endothelial Progenitor Cell Counts around the Ovulation PubMed: 22948762
Experimental & Molecular Medicine Long-term and stable correction of uremic anemia by intramuscular injection of plasmids containing hypoxia-regulated system of erythropoietin expression PubMed: PMC3509184
Biochem Biophys Res Commun. Reevaluation of erythropoietin production by the nephron Pubmed:24832733
Nutrition. Carbohydrate and glutamine supplementation modulates the Th1/Th2 balance after exercise performed at a simulated altitude of 4500 m. Pubmed:2528040
J Appl Physiol (1985). Decreased plasma soluble erythropoietin receptor in high-altitude excessive erythrocytosis and Chronic Mountain Sickness Pubmed:25324511
Expert Rev Proteomics Plasma hepcidin in early-stage breast cancer patients: no relationship with interleukin-6, erythropoietin and erythroferrone PubMed: 26496240
Oncotarget KIAA0101 is associated with human renal cell carcinoma proliferation and migration induced by erythropoietin pubmed:26575329
international journal of molecular sciences Expression of Iron-Related Proteins Differentiate Non-Cancerous and Cancerous Breast Tumors. pubmed:28216608
Veterinary Clinical Pathology Occult gastrointestinal bleeding is a common finding in dogs with chronic kidney disease pubmed:28186645
IOS Press Content Library An Intranasal Formulation of Erythropoietin (Neuro-EPO) Prevents Memory Deficits and Amyloid Toxicity in the APPSwe Transgenic Mouse Model of Alzheimer's … articles:journal-of-alzheimers-disease
Oxidative Medicine and Cellular Longevity Acidic Polysaccharide from Angelica sinensis Reverses Anemia of Chronic Disease Involving the Suppression of Inflammatory Hepcidin and NF-κB Activation pubmed:29147463
PLoS Neglected Tropical Diseases Functional and phenotypic evaluation of eosinophils from patients with the acute form of paracoccidioidomycosis pubmed:28489854
Neurochemistry International Effects of erythropoietin on astrocytes and brain endothelial cells in primary culture during anoxia depend on simultaneous signaling by other cytokines and on duration of anoxia S0197018617304783
Pesquisa Veterinária Brasileira Relation between anaemia and bone marrow features and serum erythropoietin in dogs with chronic kidney disease 10.1590/s0100-736x2017000600011 
Medical Journal Armed Forces India Study of acute hypoxia markers in healthy subjects: Utility in post-crash investigation 10.1016/j.mjafi.2017.04.003
Neurochemistry International Effects of erythropoietin on astrocytes and brain endothelial cells in primary culture during anoxia depend on simultaneous signaling by other cytokines and on … Pubmed:29180303
Clinical and Experimental Medicine Increased NGAL level associated with iron store in chronic kidney disease with anemia Pubmed:29909502
Bulletin of Experimental Biology and Medicine Signal Mechanism of the Protective Effect of Combined Preconditioning by Amtizole and Moderate Hypoxia Pubmed:29308565
Bulletin of Experimental Biology and Medicine Expression of Hif-1α, Nf-κb, and Vegf Genes in the Liver and Blood Serum Levels of HIF-1α, Erythropoietin, VEGF, TGF-β, 8-Isoprostane, and Corticosterone in Wistar Rats with High and Low Resistance to Hypoxia. Corticosterone in … Pubmed: 30353332
FASEB JOURNAL High-altitude hypoxia decreases plasma erythropoietin soluble receptor (sEpoR) concentration in humans
Journal of Functional Foods Dietary gelatin enhances non-heme iron absorption possibly via regulation of systemic iron homeostasis in rats
博士论文 PHYSIOLOGICAL ADAPTATIONS TO HEAT ACCLIMATION; REPERCUSSIONS ON CYCLING PERFORMANCE
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