Human EPCR DuoSet ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human EPCR. 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 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
Kit Content
- 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)
Scientific Data
Product Datasheets
Preparation and Storage
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.
Assay Procedure
GENERAL ELISA PROTOCOL
Plate Preparation
- Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
- Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
- Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
- Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.
Assay Procedure
- Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
- Repeat the aspiration/wash as in step 2 of Plate Preparation.
- Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Repeat the aspiration/wash as in step 2.
- Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
- Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
- Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.
Citations for Human EPCR DuoSet ELISA
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.
8
Citations: Showing 1 - 8
Filter your results:
Filter by:
-
Dysregulated Coagulation and Fibrinolysis Are Present in Patients Admitted to the Emergency Department with Acute Hypoxemic Respiratory Failure: A Prospective Study
Authors: Keskinidou, C;Vassiliou, AG;Papoutsi, E;Jahaj, E;Dimopoulou, I;Siempos, I;Kotanidou, A;
Biomedicines
Species: Human
Sample Types: Plasma
-
Lung Ultrasound Findings and Endothelial Perturbation in a COVID-19 Low-Intensity Care Unit
Authors: R Gualtierot, F Tafuri, R Rossio, M Rota, P Bucciarell, B Ferrari, A Giachi, C Suffritti, M Cugno, F Peyvandi, On Behalf
Journal of Clinical Medicine, 2022-09-15;11(18):.
Species: Human
Sample Types: Plasma
-
Procoagulant activity in high grade serous ovarian cancer patients following neoadjuvant chemotherapy-The role of the activated protein C pathway
Authors: MP Ward, FA Saadeh, SA O'Toole, JJ O'Leary, N Gleeson, LA Norris
Thrombosis Research, 2021-01-26;200(0):91-98.
Species: Human
Sample Types: Plasma
-
Expression profiles of the internal jugular and saphenous veins: Focus on hemostasis genes
Authors: N Ziliotto, S Meneghetti, E Menegatti, M Baroni, B Lunghi, F Salvi, M Ferracin, A Branchini, D Gemmati, F Mascoli, P Zamboni, F Bernardi, G Marchetti
Thromb. Res., 2020-05-03;191(0):113-124.
Species: Human
Sample Types: Plasma
-
An ADAM-10 dependent EPCR shedding links meningococcal interaction with endothelial cells to purpura fulminans
Authors: H Lécuyer, Z Virion, JP Barnier, S Matczak, S Bourdoulou, E Bianchini, F Saller, D Borgel, X Nassif, M Coureuil
PLoS Pathog., 2018-04-09;14(4):e1006981.
Species: Human
Sample Types: Cell Culture Supernates
-
The H3 Haplotype of the EPCR Gene Determines High sEPCR Levels in Critically Ill Septic Patients
Authors: AG Vassiliou, A Kotanidou, Z Mastora, C Tascini, G Cardinali, SE Orfanos
Infect Dis Ther, 2018-03-16;7(0):3-14.
Species: Human
Sample Types: Plasma
-
Elderly trauma patients have high circulating noradrenaline levels but attenuated release of adrenaline, platelets, and leukocytes in response to increasing injury severity.
Authors: Johansson PI, Sorensen AM, Perner A, Welling KL, Wanscher M, Larsen CF, Ostrowski SR
Crit. Care Med., 2012-06-01;40(6):1844-50.
Species: Human
Sample Types: Plasma
-
High sCD40L levels early after trauma are associated with enhanced shock, sympathoadrenal activation, tissue and endothelial damage, coagulopathy and mortality.
Authors: Johansson PI, Sorensen AM, Perner A, Welling KL, Wanscher M, Larsen CF, Ostrowski SR
J. Thromb. Haemost., 2012-02-01;10(2):207-16.
Species: Human
Sample Types: Plasma
FAQs
No product specific FAQs exist for this product, however you may
View all ELISA FAQsReviews for Human EPCR DuoSet ELISA
Average Rating: 5 (Based on 1 Review)
Have you used Human EPCR DuoSet ELISA?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image
Filter by:
