ELISA Kit for Erythropoietin (EPO)

EP; Epoetin; Erythropoetin; Hematopoietin; Hemopoietin

Specificity

This assay has high sensitivity and excellent specificity for detection of Erythropoietin (EPO).
No significant cross-reactivity or interference between Erythropoietin (EPO) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Erythropoietin (EPO) and the recovery rates were calculated by comparing the measured value to the expected amount of Erythropoietin (EPO) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 93-102 98
EDTA plasma(n=5) 96-105 101
heparin plasma(n=5) 97-105 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Erythropoietin (EPO) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Erythropoietin (EPO) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Erythropoietin (EPO) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 80-96% 87-101% 95-103% 93-101%
EDTA plasma(n=5) 97-105% 90-99% 85-104% 90-99%
heparin plasma(n=5) 81-101% 92-105% 78-104% 80-95%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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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
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