Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit

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DTHBD0
Control Products Available
Human Thrombomodulin/BDCA-3 ELISA Standard Curve
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Product Details
Procedure
Citations (8)
FAQs
Supplemental Products
Reviews (1)

Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (25 uL), Cell Lysates (25 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Citrate Plasma (10 uL), Urine (10 uL)
Sensitivity
27 pg/mL
Assay Range
62.5 - 4,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine)
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.

Product Summary

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.

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, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 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

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.
Human Thrombomodulin/BDCA-3 ELISA Linearity

Data Examples

Human Thrombomodulin/BDCA-3 ELISA Standard Curve

Product Datasheets

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 2 - 8 °C. Do not use past expiration date.

Background: Thrombomodulin/BDCA-3

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.

Entrez Gene IDs:
7056 (Human); 21824 (Mouse); 83580 (Rat)
Alternate Names:
BDCA3; BDCA-3; CD141 antigen; CD141; Fetomodulin; THBD; THRM; Thrombomodulin; TM
⚠ 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.

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. 100 µL Assay Diluent
  4.   Add 100 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.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.

Citations for Human Thrombomodulin/BDCA-3 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.

8 Citations: Showing 1 - 8
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  1. Permissive microbiome characterizes human subjects with a neurovascular disease cavernous angioma
    Authors: SP Polster, A Sharma, C Tanes, AT Tang, P Mericko, Y Cao, J Carrión-Pe, R Girard, J Koskimäki, D Zhang, A Stadnik, SG Romanos, SB Lyne, R Shenkar, K Yan, C Lee, A Akers, L Morrison, M Robinson, A Zafar, K Bittinger, H Kim, JA Gilbert, ML Kahn, L Shen, IA Awad
    Nat Commun, 2020;11(1):2659.
    Species: Human
    Sample Types: Plasma
  2. Enhanced effect of recombinant human soluble thrombomodulin by ultrasound irradiation in acute liver failure
    Authors: K Hoshino, Y Nakamura, T Nakano, A Watanabe, H Sheng, K Tachibana, H Ishikura
    Sci Rep, 2020;10(1):1742.
    Species: Mouse
    Sample Types: Tissue Homogenates
  3. Relationship of mottling score, skin microcirculatory perfusion indices and biomarkers of endothelial dysfunction in patients with septic shock: an observational study
    Authors: S Kazune, A Caica, K Volceka, O Suba, U Rubins, A Grabovskis
    Crit Care, 2019;23(1):311.
    Species: Human
    Sample Types: Plasma
  4. Systemic Inflammation Precedes Microalbuminuria in Diabetes
    Authors: FG Scurt, J Menne, S Brandt, A Bernhardt, PR Mertens, H Haller, C Chatzikyrk,
    Kidney Int Rep, 2019;4(10):1373-1386.
    Species: Human
    Sample Types: Serum
  5. Markers of endothelial cell activation and neutrophil extracellular traps are elevated in immune thrombocytopenia but are not enhanced by thrombopoietin receptor agonists
    Authors: L Garabet, CE Henriksson, ML Lozano, W Ghanima, J Bussel, E Brodin, MP Fernández-, C Martínez, R González-C, MC Mowinckel, PM Sandset
    Thromb. Res., 2019;185(0):119-124.
    Species: Human
    Sample Types: Plasma
  6. Effects of Alloantibodies to Human Leukocyte Antigen on Endothelial Expression and Serum Levels of Thrombomodulin
    Authors: S Béland, P Vallin, O Désy, E Lévesque, SA De Serres
    J. Thromb. Haemost, 2017;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  7. Plasma irisin is elevated in type 2 diabetes and is associated with increased E-selectin levels
    Authors: KS Rana, C Pararasa, I Afzal, DA Nagel, EJ Hill, CJ Bailey, HR Griffiths, I Kyrou, HS Randeva, S Bellary, JE Brown
    Cardiovasc Diabetol, 2017;16(1):147.
    Species: Human
    Sample Types: Plasma
  8. Common genetic risk factors for venous thrombosis in the chinese population.
    Am J Hum Genet, 2013;92(2):177-87.
    Species: Human
    Sample Types: Plasma

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Human Thrombomodulin/BDCA-3 Quantikine ELISA Kit
By Anonymous on 07/18/2017
Application: Sample Tested: Citrate Plasma