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Mouse Erythropoietin/EPO Quantikine ELISA Kit Best Seller

Bio-Techne includes R&D Systems | Catalog # MEP00B

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MEP00B

Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Tissue Homogenates (50 µL), Serum (25 µL), Heparin Plasma (25 µL)

Sensitivity

46.9 pg/mL

Assay Range

47-3000 pg/mL (Cell Culture Supernates, Tissue Homogenates, Serum, Heparin Plasma)

Product Summary for Mouse Erythropoietin/EPO Quantikine ELISA Kit

The Quantikine Mouse Epo Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse Epo in cell culture supernates, tissue homogenates, serum, and plasma. It contains antibodies raised against recombinant Epo and has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse Epo showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that the Quantikine Mouse Epo kit can be used to determine relative mass values for naturally occurring mouse Epo.

Product Specifications

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Mouse

Specificity

Natural and recombinant mouse Epo. Recombinant rat Epo has 100% cross-reactivity in this kit. This kit is not validated for use with rat samples, because rat sample dilution is not linear.

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

Interference observed with 1 or more available related molecules.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates, Heparin Plasma, Serum, Tissue Homogenates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 196 258 704 180 239 700
Standard Deviation 7.7 7.8 31.2 17.5 15.3 17.5
CV% 3.9 3.0 4.4 9.7 6.4 2.5

Recovery for Mouse Erythropoietin/EPO Quantikine ELISA Kit

The recovery of Epo spiked to three levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=7) 110 102-117
Heparin Plasma (n=8) 103 90-120
Serum (n=9) 93 85-114
Tissue Homogenates (n=3) 94 80-111

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of Epo in each matrix were diluted with Calibrator Diluent and then assayed.

Mouse Erythropoietin ELISA Linearity

Scientific Data Images for Mouse Erythropoietin/EPO Quantikine ELISA Kit

Mouse Erythropoietin ELISA Standard Curve

Mouse Erythropoietin ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Erythropoietin/EPO

Erythropoietin (Epo) is a 34-39 kDa secreted glycoprotein that is a member of the type I cytokine superfamily. The mouse Epo gene encodes a 192 amino acid (aa) residue precursor that contains a 26 aa signal peptide and a 166 aa mature protein containing three potential N-linked glycosylation sites (1-4). Mouse Epo lacks the O-linked glycosylation site found in human Epo. Although carbohydrate chains are not required for in vitro receptor binding, they are required for in vivo Epo bioactivity. Depending on the cell source, different Epo isoforms are produced that differ in their glycan compositions and sialic acid contents (5-8). Mature mouse and rat Epo share 94% aa sequence identity. They also share from 80%-82% aa identity with mature human, porcine, rhesus monkey and feline Epo (2, 3). Epo is primarily produced by cells in the kidney (interstitial peritubular renal fibroblasts) and liver (hepatocytes and Ito cells), where its production is up-regulated by hypoxia. Other tissues and cells, including neural tissues (astrocytes and neurons), testis (Sertoli cells), uterus, placenta, and erythroid progenitors, have also been shown to produce Epo (9-14). 
Epo is best known for its role in red blood cell formation. While Epo is not a lineage commitment factor, it inhibits apoptosis and induces burst forming unit-erythroid (BFU-E) differentiation into colony forming unit-erythroid (CFU-E), and the subsequent proliferation and maturation of CFU-E into early normoblasts (10, 15, 16). Apart from its role in erythropoiesis, Epo also acts on various non-hematopoietic cells to function as a viability and proliferation factor. Epo can stimulate myoblast proliferation while suppressing its differentiation, resulting in the expansion of the progenitor cell population (17). Epo is a tissueprotective factor that protects against ischemic and toxic injuries to neuronal, cardiovascular and renal tissues (18, 19). Epo has also been shown to promote angiogenesis in various physiologic and pathologic conditions (20, 21). 
Epo binds and signals via the high-affinity preformed homodimeric Epo receptor (Epo R) that is composed of two Epo R subunits. Each Epo R subunit is a type I transmembrane glycoprotein that belongs to the type I cytokine receptor superfamily (18, 22-24). Its extracellular domain contains the characteristic two fibronectin type III domains and a WSxWS motif near the plasma membrane (24, 25). Binding of Epo to the Epo R homodimer results in conformational change and phosphorylation and activation of the non-receptor protein kinase JAK2, which activates the downstream signaling cascade (26). An alternative Epo heteromeric receptor complex that transduces cell-protective signals and containing the beta common receptor ( betaCR) subunit in addition to the Epo R subunit has been described. betaCR also belongs to the type I cytokine receptor superfamily and is a subunit that is shared by the heteromeric IL-3, IL-5 and GM-CSF receptor complexes. Epo binds with lower affinity to the heteromeric receptor consisting of a Epo R subunit and a betaCR homodimer (18).

Alternate Names

ECYT5, EPO, MVCD2

Entrez Gene IDs

2056 (Human); 13856 (Mouse); 24335 (Rat)

Gene Symbol

EPO

Additional Erythropoietin/EPO Products

Product Documents for Mouse Erythropoietin/EPO Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse Erythropoietin/EPO Quantikine ELISA Kit

For research use only

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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.