Mouse Thrombomodulin/BDCA-3 Quantikine ELISA Kit Summary
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine
|Intra-Assay Precision||Inter-Assay Precision|
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|
Preparation and Storage
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.
- 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 50 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 100 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 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- 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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