Mouse Thrombomodulin/BDCA-3 Quantikine ELISA Kit

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MTHBD0
Mouse Thrombomodulin/BDCA-3 Standard Curve
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Mouse 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 (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Urine (10 uL)
Sensitivity
19.1 pg/mL
Assay Range
46.9 - 3,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
Specificity
Natural and recombinant mouse Thrombomodulin.
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 Mouse Thrombomodulin/BDCA-3 immunoassay is a 4.5 hour solid-phase ELISA designed to measure Thrombomodulin in mouse cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant mouse Thrombomodulin and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant mouse Thrombomodulin. Results obtained using natural mouse Thrombomodulin showed dose response curves that were parallel to the standard curves obtained using the Quantikine mouse kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse Thrombomodulin.

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 twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of kit components

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 108 388 1159 121 380 1155
Standard Deviation 6.67 16 44.8 16 38.1 136
CV% 6.2 4.1 3.9 13.2 10 11.8

Recovery

The recovery of mouse Thrombomodulin spiked to levels throughout the range of the assay was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 104 91-115

Linearity

To assess the linearity of the assay, samples containing high concentrations of mouse Thrombomodulin in each matrix were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay. All samples were diluted prior to assay.
Mouse Thrombomodulin/BDCA-3 ELISA Linearity

Product Datasheets

Preparation and Storage

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. 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.
  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.
  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. 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.

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