Human EPCR Quantikine ELISA Kit

Discontinued Product

DEPCR0 has been discontinued.
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Human EPCR Quantikine ELISA Kit Summary

Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (20 uL), EDTA Plasma (20 uL), Heparin Plasma (20 uL)
Sensitivity
0.282 ng/mL
Assay Range
0.6 - 40 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human EPCR
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human EPCR Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human EPCR in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant human EPCR and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human EPCR showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring EPCR.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times 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

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 5.2 11.7 21.5 5.26 10.7 21
Standard Deviation 0.2 0.6 1.7 0.3 0.7 1.3
CV% 4.8 4.9 7.7 5.4 6.4 6

Recovery

The recovery of human EPCR spiked to three levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Samples (n=6) 93 86-99

Linearity

To assess the linearity of the assay, samples containing or spiked with high concentrations of EPCR were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: EPCR

EPCR (the Endothelial Protein C Receptor), also known as CD201, is a transmembrane glyoprotein expressed on vascular endothelial cells. EPCR inhibits thrombosis through its interactions with Protein C, activated Protein C (APC), and Coagulation Factors VII, and VIIa. It enhances the activation of Protein C in response to complexes of Thrombin-Thrombomodulin. A soluble form of EPCR inhibits the anti-coagulant activity of APC. EPCR binds to CD11b/CD18 (Mac-1) on monocytes and mediates monocyte adhesion to the vascular endothelium. In addition, it binds to the antigen receptor on gamma/delta T cells, promotes hematopoietic stem cell retention in the bone marrow, and contributes to malaria pathogenicity through binding surface proteins on Plasmodium.

Long Name:
Endothelial Protein C Receptor
Entrez Gene IDs:
10544 (Human); 19124 (Mouse)
Alternate Names:
APC receptor; CCCA; CCD41; CCD41centrosome-associated protein; CD201 antigen; CD201; centrocyclin; Endothelial cell protein C receptor; endothelial protein C receptor; EPCR; EPCRMGC23024; PROCR; protein C receptor, endothelial

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 100 µL Conjugate
  9.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
  10.   Aspirate and wash 4 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citation for Human EPCR Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

  1. Cellular regulation of blood coagulation: a model for venous stasis.
    Authors: Campbell JE, Brummel-Ziedins KE, Butenas S
    Blood, 2010-09-23;116(26):6082-91.
    Species: Human
    Sample Types: Cell Lysates

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