Thrombomodulin (TM), also known as CD141 and BDCA3, is a transmembrane expressed on vascular endothelial cells (EC), arterial smooth muscle cells, monocytes, and macrophages. It binds Thrombin and enhances the Thrombin-mediated activation of the anti-coagulant Protein C and the anti-fibrinolytic TAFI/Carboxypeptidase B2. Thrombomodulin also inhibits the ability of Thrombin to activate several pro-coagulant proteins (e.g. Fibrinogen, Factor V, Factor XIII, PAR-1). Thrombomodulin exerts multiple anti-inflammatory effects and reduces the severity of disease involving chronic inflammation. In cancer, Thrombomodulin suppresses tumor angiogenesis, growth, and epithelial-mesenchymal transition. Soluble fragments of Thrombomodulin are elevated in the serum, urine, and synovial fluid during coagulation disorders, inflammation, and organ failure.
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Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit
R&D Systems | Catalog # DTHBD0
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (25 µL), Cell Lysates (25 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL), Citrate Plasma (10 µL), Urine (10 µL)
Sensitivity
27 pg/mL
Assay Range
62.5-4000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
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Product Summary for Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit
The Quantikine Human Thrombomodulin Immunoassay is a 4.5 hour solid phase ELISA designed to measure human Thrombomodulin in cell culture supernates, cell lysates, serum, plasma, and urine. It contains NS0-expressed recombinant human Thrombomodulin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Thrombomodulin 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 human Thrombomodulin.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Human
Specificity
Natural and recombinant human Thrombomodulin. This kit also recognizes Thrombomodulin complexed with Thrombin, Protein C, and Thrombin/Protein C.
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.
精度
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, Cell Lysates, Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 571 | 1123 | 2318 | 518 | 1073 | 2159 |
| Standard Deviation | 13.2 | 31.1 | 83.9 | 41.2 | 76.1 | 123 |
| CV% | 2.3 | 2.8 | 3.6 | 7.9 | 7.1 | 5.7 |
Recovery for Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit
The recovery of Thrombomodulin spiked to levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 96 | 88-106 |
| Citrate Plasma (n=4) | 94 | 85-102 |
| EDTA Plasma (n=4) | 95 | 86-105 |
| Heparin Plasma (n=4) | 91 | 85-96 |
| Serum (n=4) | 89 | 85-96 |
| Urine (n=4) | 99 | 85-115 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of Thrombomodulin were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit
Human Thrombomodulin/BDCA-3 ELISA Standard Curve
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: Thrombomodulin/BDCA-3
Alternate Names
BDCA-3, BDCA3, CD141, Fetomodulin, THBD, THRM
Gene Symbol
THBD
Additional Thrombomodulin/BDCA-3 Products
Product Documents for Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit
Product Specific Notices for Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit
For research use only
⚠ 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.Citations for Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit
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Protocols
View specific protocols for Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit (DTHBD0):
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.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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