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Mouse Erythropoietin/EPO DuoSet ELISA

Bio-Techne includes R&D Systems | Catalog # DY959

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DY959

Ancillary Products Available

Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

62.5-4000 pg/mL

Sample Type

Cell culture supernates, serum, and plasma
Note: Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet

Reactivity

Mouse

Mouse Erythropoietin/EPO DuoSet ELISA Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Product Summary for Mouse Erythropoietin/EPO DuoSet ELISA

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse Erythropoietin. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

Product Specifications

Assay Format

96-well strip plate (sold separately)

Sample Volume Required

100 µL

Detection Method

Colorimetric ELISA - 450nm (TMB)

Conjugate

Biotin

Label

HRP

Scientific Data Images for Mouse Erythropoietin/EPO DuoSet ELISA

Mouse Erythropoietin ELISA Standard Curve

Mouse Erythropoietin ELISA Standard Curve

Kit Contents for Mouse Erythropoietin/EPO DuoSet ELISA

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Preparation and Storage

Shipping

The product is shipped at ambient temperature. 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 DuoSet ELISA

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 DuoSet ELISA

For research use only

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