Human sCD141 ELISA Assay


The Human sCD141 ELISA Assay kit is a solid phase sandwich ELISA for the in-vitro qualitative and quantitative determination of sCD141 in supernatants, buffered solutions or serum and plasma samples. This assay will recognise both natural and recombinant human sCD141. The Eagle Biosciences Human sCD141 ELISA Assay kit is for Research Use Only.

SKU: C4131-K01 Categories: , ,

Human sCD141 ELISA Assay

The Human sCD141 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.31 ng/ml
Dynamic Range: 0.625 – 20 ng/ml
Incubation Time: 1h45mn
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Name: Thrombomodulin, BDC-A

Assay Background

CD141 , also known as Thrombomodulin or BDC-A is an transmembrane protein expressed predominantly on the surface of vascular endothelial cells, but also on other cell types such as macrophages, monocytes, subpopulation of myeloid dendritic cells and keratinocytes. Thrombomodulin is a type 1 membrane glycoprotein of 74kDa consisting on a single chain with 6 Epidermal Growth Factor (EGF)-like domains critical for Thrombin and proteine C binding, and four other domains (an N-terminal C-type lectin domain, a highly glycosylated serine/threonine-rich domain, a transmembrane segment, and a short C-terminal cytoplasmic tail. It acts as an important component in the anti-coagulation and fibrinolysis system : cofactor for active thrombin, the thrombomodulin-thrombin complex activates protein C to degrade coagulation factors, inhibiting coagulation and fibrinolysis.

When vascular endothelial cells injury occurs, proteolytic cleavage release multiple fragments of thrombomodulin in circulation, so it represents a good marker for endothelial cell damage in pathologies such as cardiovascular disease, acute coronary syndrome, pulmonary thromboembolism or severe hemorrhage. Thrombomodulin has been described as an anticoagulant effect protein, and its therapeutical potential for preventing deep vein thrombosis and its use in the treatment of disseminated intravascular coagulation is under demonstrated in preliminary data.

More recently, other biological effects beyond anticoagulation have been demonstrated such as an anti-inflammatory , cell adhesion and proliferation regulation It is also demonstrated that thrombomodulin may be able to suppress the metastatic capacity of tumor cells, and hence have a protective role in many types of malignancies.

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Additional Information

Assay Principle

A capture Antibody highly specific for CD141 has been coated to the wells of the microtitre strip plate provided during manufacture. Binding of CD141 samples and known standards to the capture antibodies and subsequent binding of the biotinylated anti-CD141 secondary antibody to the analyte is completed during the same incubation period. Any excess unbound analyte and secondary antibody is removed. The HRP conjugate solution is then added to every well including the zero wells, following incubation excess conjugate is removed by careful washing. A chromogen substrate is added to the wells resulting in the progressive development of a blue coloured complex with the conjugate. The colour development is then stopped by the addition of acid turning the resultant final product yellow. The intensity of the produced coloured complex is directly proportional to the concentration of CD141 present in the samples and standards. The absorbance of the colour complex is then measured and the generated OD values for each standard are plotted against expected concentration forming a standard curve. This standard curve can then be used to accurately determine the concentration of CD141 in any sample tested.

  1. Add 100μl of each, Sample, Standard, Control and zero ( standard diluent) in duplicate to appropriate number of wells
  2. Add 50μl of diluted biotinylated anti-CD141 to all wells
  3. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour(s)
  4. Remove the cover and wash the plate
  5. Add 100μl of Streptavidin-HRP solution into all wells
  6. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  7. Repeat wash step 5.
  8. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  9. Incubate in the dark for 10-15 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil.
  10. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve

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